human desaturase gene and uses thereof

ABSTRACT

The subject invention relates to the identification of a gene involved in the desaturation of polyunsaturated fatty acids at carbon 5 (i.e., “human Δ5-desaturase”) and to uses thereof. In particular, human Δ5-desaturase may be utilized, for example, in the conversion of dihomo-γ-linolenic acid (DGLA) to arachidonic acid (AA) and in the conversion of 20:4n−3 to eicosapentaenoic acid (EPA). AA or polyunsaturated fatty acids produced therefrom may be added to pharmaceutical compositions, nutritional compositions, animal feeds, as well as other products such as cosmetics.

[0001] The subject application is a Continuation-In-Part of pendingInternational Application PCT/US98/07422 filed on Apr. 10, 1998 (whichdesignates the U.S.) which is a Continuation-In-Part of pending U.S.patent application Ser. No. 08/833,610 filed on Apr. 11, 1997.

BACKGROUND OF THE INVENTION

[0002] 1. Technical Field

[0003] The subject invention relates to the identification and isolationof a gene that encodes an enzyme (i.e., human Δ5-desaturase) involved inthe synthesis of polyunsaturated fatty acids and to uses thereof. Inparticular, Δ5-desaturase catalyzes the conversion of, for example,dihomo-y-linolenic acid (DGLA) to arachidonic acid (AA) and(n−3)-eicosatetraenoic acid (20:4n−3) to eicosapentaenoic acid(20:5n−3). The converted product may then be utilized as a substrate inthe production of other polyunsaturated fatty acids (PUFAs). The productor other polyunsaturated fatty acids may be added to pharmaceuticalcompositions, nutritional composition, animal feeds as well as otherproducts such as cosmetics.

[0004] 2. Background Information

[0005] Desaturases are critical in the production of long-chainpolyunsaturated fatty acids which have many important functions. Forexample, PUFAs are important components of the plasma membrane of acell, where they are found in the form of phospholipids. They also serveas precursors to mammalian prostacyclins, eicosanoids, leukotrienes andprostaglandins. Additionally, PUFAs are necessary for the properdevelopment of the developing infant brain as well as for tissueformation and repair. In view of the biological significance of PUFAs,attempts are being made to produce them, as well as intermediatesleading to their production, in an efficient manner.

[0006] A number of enzymes are involved in PUFA biosynthesis includingΔ5-desaturase (see FIG. 11). For example, elongase (elo) catalyzes theconversion of γ-linolenic acid (GLA) to dihomo-γ-linolenic acid (DGLA)and of stearidonic acid (18:4n−3) to (n−3)-eicosatetraenoic acid(20:4n−3). Linoleic acid (LA, 18:2−Δ9, 12 or 18:2n−6) is produced fromoleic acid (18:1−Δ9) by a Δ12-desaturase. GLA (18:3−Δ6, 9, 12) isproduced from linoleic acid by a Δ6-desaturase.

[0007] It must be noted that animals cannot desaturate beyond the Δ9position and therefore cannot convert oleic acid into linoleic acid.Likewise, α-linolenic acid (ALA, 18:3−Δ9, 12, 15) cannot be synthesizedby mammals. However, α-linolenic acid can be converted to stearidonicacid (STA, 18:4−Δ6, 9, 12, 15) by a Δ6-desaturase (see PCT publicationWO 96/13591 and The Faseb Journal, Abstracts, Part I, Abstract 3093,page A532 (Experimental Biology 98, San Francisco, Calif., Apr. 18-22,1998) see also U.S. Pat. No. 5,552,306), followed by elongation to(n−3)-eicosatetraenoic acid (20:4−Δ8, 11, 14, 17) in mammals and algae.This polyunsaturated fatty acid (i.e., 20:4−Δ8, 11, 14, 17) can then beconverted to eicosapentaenoic acid (EPA, 20:5−Δ5, 8, 11, 14, 17) by aΔ5-desaturase, such as that of the present invention. Other eukaryotes,including fungi and plants, have enzymes which desaturate at carbon 12(see PCT publication WO 94/11516 and U.S. Pat. No. 5,443,974) and carbon15 (see PCT publication WO 93/11245). The major polyunsaturated fattyacid of animals therefore are either derived from diet and/or fromdesaturation and elongation of linoleic acid or α-linolenic acid. Inview of these difficulties, it is of significant interest to isolategenes involved in PUFA synthesis from species that naturally producethese fatty acids and to express these genes in a microbial, plant, oranimal system which can be altered to provide production of commercialquantities of one or more PUFAs. One of the most important long chainPUFAs, noted above, is arachidonic acid (AA). AA is found in filamentousfungi and can also be purified from mammalian tissues including theliver and adrenal glands. As noted above, AA production fromdihomo-γ-linolenic acid is catalyzed by a Δ5-desaturase. EPA is anotherimportant long-chain PUFA. EPA is found in fungi and also in marineoils. As noted above, EPA is produced from (n−3)-eicosatetraenoic acidand is catalyzed by a Δ5-desaturase.

[0008] In view of the above discussion, there is a definite need for theΔ5-desaturase enzyme, the gene encoding this enzyme, as well asrecombinant methods of producing this enzyme. Additionally, a needexists for oils containing levels of PUFAs beyond those naturallypresent as well as those enriched in novel PUFAs. Such oils can only bemade by isolation and expression of the Δ5-desaturase gene.

[0009] All U.S. patents and publications referred to herein are herebyincorporated in their entirety by reference.

SUMMARY OF THE INVENTION

[0010] The present invention includes an isolated nucleotide sequencecorresponding to or complementary to at least about 50% of thenucleotide sequence shown in SEQ ID NO:1 (FIG. 12).

[0011] The isolated nucleotide sequence may be represented by SEQ IDNO:1. These sequences may encode a functionally active desaturase whichutilizes a polyunsaturated fatty acid as a substrate. The sequences maybe derived from a mammal such as, for example, a human.

[0012] The present invention also includes purified proteins encoded bythe nucleotide sequences referred to above. Additionally, the presentinvention includes a purified polypeptide which desaturatespolyunsaturated fatty acids at carbon 5 and has at least about 50% aminoacid similarity to the amino acid sequence of the purified proteinsreferred to directly above.

[0013] Furthermore, the present invention also encompasses a method ofproducing a human Δ5-desaturase. This method comprises the steps of: a)isolating the nucleotide sequence represented by SEQ ID NO:1 (FIG. 12);b) constructing a vector comprising: i) the isolated nucleotide sequenceoperably linked to ii) a promoter; and c) introducing the vector into ahost cell under time and conditions sufficient for expression of thehuman Δ5-desaturase. The host cell may be, for example, a eukaryoticcell or a prokaryotic cell. In particular, the prokaryotic cell may be,for example, E. coli, cyanobacteria or B. subtilis. The eukaryotic cellmay be, for example, a mammalian cell, an insect cell, a plant cell or afungal cell (e.g., a yeast cell such as Saccharomyces cerevisiae,Saccharomyces carlsbergensis, Candida spp., Lipomyces starkey, Yarrowialipolytica, Kluyveromyces spp., Hansenula spp., Trichoderma spp. orPichia spp.).

[0014] Additionally, the present invention also encompasses a vectorcomprising: a) a nucleotide sequence as represented by SEQ ID NO:1 (FIG.12) operably linked to b) a promoter. The invention also includes a hostcell comprising this vector.

[0015] The host cell may be, for example, a eukaryotic cell or aprokaryotic cell. Suitable eukaryotic cells and prokaryotic cells are asdefined above.

[0016] Moreover, the present invention also includes a plant cell, plantor plant tissue comprising the above vector, wherein expression of thenucleotide sequence of the vector results in production of apolyunsaturated fatty acid by the plant cell, plant or plant tissue. Thepolyunsaturated fatty acid may be, for example, selected from the groupconsisting of AA and EPA. The invention also includes one or more plantoils or acids expressed by the above plant cell, plant or plant tissue.

[0017] Additionally, the present invention also encompasses a transgenicplant comprising the above vector, wherein expression of the nucleotidesequence of the vector results in production of a polyunsaturated fattyacid in seeds of the transgenic plant.

[0018] Also, the invention includes a mammalian cell comprising theabove vector wherein expression of the nucleotide sequence of the vectorresults in production of altered levels of AA or EPA when the cell isgrown in a culture media comprising a fatty acid selected from the groupconsisting of an essential fatty acid, LA and ALA.

[0019] It should also be noted that the present invention encompasses atransgenic, non-human mammal whose genome comprises a DNA sequenceencoding a human Δ5-desaturase operably linked to a promoter. The DNAsequence may be represented by SEQ ID NO:1 (FIG. 12). Additionally, thepresent invention includes a fluid (e.g., milk) produced by thetransgenic, non-human mammal wherein the fluid comprises a detectablelevel of at least human Δ5-desaturase.

[0020] Additionally, the present invention includes a method (i.e.,“first” method) for producing a polyunsaturated fatty acid comprisingthe steps of: a) isolating the nucleotide sequence represented by SEQ IDNO:1 (FIG. 12); b) constructing a vector comprising the isolatednucleotide sequence; c) introducing the vector into a host cell undertime and conditions sufficient for expression of the human Δ5-desaturaseenzyme; and d) exposing the expressed human Δ5-desaturase enzyme to asubstrate polyunsaturated fatty acid in order to convert the substrateto a product polyunsaturated fatty acid. The substrate polyunsaturatedfatty acid may be, for example, DGLA or 20:4n−3 and the productpolyunsaturated fatty acid may be, for example, AA or EPA, respectively.This method may further comprise the step of exposing the productpolyunsaturated fatty acid to an elongase in order to convert theproduct polyunsaturated fatty acid to another polyunsaturated fatty acid(i.e., “second” method). In this method containing the additional step(i.e., “second” method), the product polyunsaturated fatty acid may be,for example, AA or EPA, and the “another” polyunsaturated fatty acid maybe adrenic acid or (n−3)-docosapentaenoic acid, respectively. The methodcontaining the additional step may further comprise a step of exposingthe another polyunsaturated fatty acid to an additional desaturase inorder to convert the another polyunsaturated fatty acid to a finalpolyunsaturated fatty acid (i.e., “third” method). The finalpolyunsaturated fatty acid may be, for example, (n−6)-docosapentaenoicacid or docosahexaenoic (DHA) acid.

[0021] The present invention also encompasses a nutritional compositioncomprising at least one polyunsaturated fatty acid selected from thegroup consisting of the product polyunsaturated fatty acid producedaccording to the “first” method, another polyunsaturated fatty acidproduced according to the “second” method, and the final polyunsaturatedfatty acid produced according to the “third” method. The productpolyunsaturated fatty acid may be, for example, AA or EPA. The anotherpolyunsaturated fatty acid may be, for example, adrenic acid or(n−3)-docosapentaenoic acid. The final polyunsaturated fatty acid maybe, for example, (n−6)-docosapentaenoic acid or DHA. This nutritionalcomposition, may be, for example, an infant formula, a dietarysupplement or a dietary substitute and may be administered to a human orto an animal. It may be administered enterally or parenterally. Thenutritional composition may further comprise at least one macronutrientselected from the group consisting of coconut oil, soy oil, canola oil,monoglycerides, diglycerides, glucose, edible lactose, electrodialysedwhey, electrodialysed skim milk, milk whey, soy protein, and proteinhydrolysates. Additionally, the composition may further comprise atleast one vitamin selected from the group consisting of Vitamins A, C,D, E, and B complex and at least one mineral selected from the groupconsisting of calcium magnesium, zinc, manganese, sodium, potassium,phosphorus, copper, chloride, iodine, selenium and iron.

[0022] Furthermore, the present invention also includes a apharmaceutical composition comprising 1) at least one polyunsaturatedfatty acid selected from the group consisting of the productpolyunsaturated fatty acid produced according to the “first” method, theanother polyunsaturated fatty acid produced according to the “second”method, and the final polyunsaturated fatty acid produced according tothe “third” method and 2) a pharmaceutically acceptable carrier. Again,the pharmaceutical composition may be administered to a human or to ananimal.

[0023] The composition may further comprise an element selected from thegroup consisting of a vitamin, a mineral, a carbohydrate, an amino acid,a free fatty acid, a phospholipid, an antioxidant, and a phenoliccompound.

[0024] Additionally, the present invention includes an animal feedcomprising at least one polyunsaturated fatty acid selected from thegroup consisting of the product polyunsaturated fatty acid producedaccording to the first method, the another polyunsaturated fatty acidproduced according to the second method and the final polyunsaturatedfatty acid produced according to the third method. The productpolyunsaturated fatty acid may be, for example, AA or EPA. The anotherpolyunsaturated fatty acid may be, for example, adrenic acid or(n−3)-docosapentaenoic acid. The final polyunsaturated fatty acid maybe, for example,(n−6)-docosapentaenoic acid or DHA.

[0025] Moreover, the present invention also includes a cosmeticcomprising a polyunsaturated fatty acid selected from the groupconsisting of the product polyunsaturated fatty acid produced accordingto the first method, the another polyunsaturated fatty acid producedaccording to the second method, and the final polyunsaturated fatty acidproduced according to the third method.

[0026] Additionally, the present invention encompasses a method ofpreventing or treating a condition caused by insufficient intake ofpolyunsaturated fatty acids comprising administering to the patient thenutritional composition of above in an amount sufficient to effectprevention or treatment.

BRIEF DESCRIPTION OF THE DRAWINGS

[0027]FIG. 1 outlines the sections of the M. alpina Δ5- andΔ6-desaturases, the clone ID's from the LifeSeq database to which thosesections had homology, and the keyword associated with the clone ID's.

[0028]FIG. 2 represents the contig 2692004.

[0029]FIG. 3 represents the contig 2153526.

[0030]FIG. 4 represents the contig 3506132.

[0031]FIG. 5 represents the contig 3854933.

[0032]FIG. 6 represents the contig 2511785.

[0033]FIG. 7 represents the contig 2535 generated based on contig2511785 of FIG. 6 and contig 3506132 of FIG. 4.

[0034]FIG. 8 represents the contig 253538a generated based on contig2535 of FIG. 7 and contig 3854933 of FIG. 5.

[0035]FIG. 9 represents the amino acid sequence identity between the M.alpina Δ5-desaturase (Ma29) and the contig 253538a.

[0036]FIG. 10 represents the amino acid sequence identity between the M.alpina Δ6-desaturase (Ma524) and the contig 253538a.

[0037]FIG. 11 represents various fatty acid biosynthesis pathways. Therole of the Δ5-desaturase enzyme should be noted.

[0038]FIG. 12 represents the complete nucleotide sequence of the humanΔ5-desaturase gene (human Δ5).

[0039]FIG. 13 represents the amino acid sequence of the humanΔ5-desaturase translated from human Δ5 (see FIG. 12).

[0040]FIG. 14 illustrates the sequence identity between the pRAE-7 andpRAE-8 clones.

[0041]FIG. 15 represents the complete putative human desaturase genesequence from clone pRAE-7.

[0042]FIG. 16 illustrates the amino acid sequence identity between theputative human desaturase gene in pRAE-7 and the M. alpinaΔ5-desaturase.

[0043]FIG. 17 illustrates the amino acid sequence identity between theputative human desaturase gene in pRAE-7 and the M. alpinaΔ6-desaturase.

[0044]FIG. 18 illustrates the amino acid sequence identity between theputative human desaturase gene in pRAE-7 and the contig 2535.

[0045]FIG. 19 illustrates the amino acid sequence identity between theputative human desaturase gene in pRAE-7 and the contig 38.

[0046]FIG. 20 illustrates the amino acid sequence identity between theN-terminus of clone A-1, a representative of Group 1, and the N-terminusof cytochrome b5 gene.

[0047]FIG. 21 illustrates the nucleotide sequence identity between thenucleotide sequence of a portion of clone A-1 and a portion of theGenBank sequence ac004228.

[0048]FIG. 22 represents the nucleotide sequence identity between thenucleotide sequence of a portion of clone 3-5 of Group 2 and a portionof the GenBank sequence ac004228. Clone 3-5 has an ATG within a NcoIsite, but translates four stops between the ATG and the BamHI site.

[0049]FIG. 23 represents the nucleotide sequence identity between thenucleotide sequence of a portion of clone A-10 of Group 3 and a portionof the GenBank sequence ac004228. Clone A-10 has an ATG 135 bp upstreamof the BamHI site, giving an open reading frame of 1267 bp.

[0050]FIG. 24 represents the nucleotide sequence identity between thenucleotide sequence of a portion of clone A-16 of Group 4 and a portionof the GenBank sequence ac004228. Clone A-16 does not have an ATG;however, there is an ATG (underlined) upstream of where the sequencealigns with ac004228.

[0051]FIG. 25 represents the nucleotide sequence identity between thenucleotide sequence of a portion of clone A-19 of Group 5 and a portionof the GenBank sequence ac004228. Clone A-19 does not have an ATG;however, this clone matches the ac004228 sequence even upstream of theBamHI site.

[0052]FIG. 26 represents the partial nucleotide sequence of the GenBanksequence ac004228 and the representative clones from the five Groups.

[0053]FIG. 27 represents the nucleotide sequence identity between thehuman Δ5-desaturase and contig 3381584.

[0054]FIG. 28 represents the nucleotide sequence identity between thehuman Δ5-desaturase and contig 2153526.

[0055]FIG. 29 represents the amino acid sequence identity between thehuman Δ5-desaturase and contig 253538a.

[0056]FIG. 30 represents the amino acid sequence identity between thehuman Δ5-desaturase and contig 38.

[0057]FIG. 31 represents the amino acid sequence identity between the M.alpina Δ6-desaturase (Ma524) and the human the Δ5-desaturase.

[0058]FIG. 32 represents the amino acid sequence identity between the M.alpina Δ5-desaturase (Ma29) and the human Δ5-desaturase.

[0059]FIG. 33 illustrates the human Δ5-desaturase activity of the genein clone pRAE-28-5, compared to that in pRAE-26-1, pRAE-33, and pRAE-35,when expressed in baker's yeast.

[0060]FIG. 34 illustrates the substrate specificity of the humanΔ5-desaturase gene in clone pRAE-28-5, converting DGLA(20:3n−6) toAA(20:4n−6), when the gene is expressed in baker's yeast.

DETAILED DESCRIPTION OF THE INVENTION

[0061] The subject invention relates to the nucleotide and amino acidsequence of the Δ5-desaturase gene derived from humans. Furthermore, thesubject invention also includes uses of the gene and of the enzymeencoded by this gene. For example, the gene and corresponding enzyme maybe used in the production of polyunsaturated fatty acids such as, forinstance, arachidonic acid, eicosapentaenoic acid, and/or adrenic acidwhich may be added to pharmaceutical compositions, nutritionalcompositions and to other valuable products.

[0062] The Human Δ5-Desaturase Gene and Enzyme Encoded Thereby

[0063] As noted above, the enzyme encoded by the human Δ5-desaturasegene is essential in the production of highly unsaturatedpolyunsaturated fatty acids having a length greater than 20 carbons. Thenucleotide sequence of the isolated human Δ5-desaturase gene is shown inFIG. 2, and the amino acid sequence of the corresponding purifiedprotein is shown in FIG. 3.

[0064] As an example, the isolated human Δ5-desaturase gene of thepresent invention converts DGLA to AA or converts 20:4n−3 to EPA. Thus,neither AA nor EPA, for example, can be synthesized without theΔ5-desaturase gene (e.g., human or M. alpina) and enzyme encodedthereby.

[0065] It should be noted that the present invention also encompassesnucleotide sequences (and the corresponding encoded proteins) havingsequences corresponding to or complementary to at least about 50%,preferably at least about 60%, and more preferably at least about 70% ofthe nucleotides in sequence to SEQ ID NO:1 (i.e., the nucleotidesequence of the human Δ5-desaturase gene described herein (see FIG.12)). Such sequences may be derived from non-human sources (e.g., C.elegans or mouse). Furthermore, the present invention also encompassesfragments and derivatives of the nucleotide sequence of the presentinvention (i.e., SEQ ID NO:1), as well as of the sequences derived fromnon-human sources, and having the above-described complementarity orcorrespondence. Functional equivalents of the above-sequences (i.e.,sequences having human Δ5-desaturase activity) are also encompassed bythe present invention. The invention also includes a purifiedpolypeptide which desaturates polyunsaturated fatty acids at the carbon5 position and has at least about 50% amino acid similarity to the aminoacid sequence of the above-noted proteins which are, in turn, encoded bythe above-described nucleotide sequences.

[0066] The present invention also encompasses an isolated nucleotidesequence which encodes PUFA desaturase activity and that ishybridizable, under moderately stringent conditions, to a nucleic acidhaving a nucleotide sequence corresponding to or complementary to thenucleotide sequence represented by SEQ ID NO:1 and shown in FIG. 12. Anucleic acid molecule is “hybridizable” to another nucleic acid moleculewhen a single-stranded form of the nucleic acid molecule can anneal tothe other nucleic acid molecule under the appropriate conditions oftemperature and ionic strength (see Sambrook et al., “Molecular Cloning:A Laboratory Manual, Second Edition (1989), Cold Spring HarborLaboratory Press, Cold Spring Harbor, N.Y.)). The conditions oftemperature and ionic strength determine the “stringency” of thehybridization. “Hybridization” requires that two nucleic acids containcomplementary sequences. However, depending on the stringency of thehybridization, mismatches between bases may occur. The appropriatestringency for hybridizing nucleic acids depends on the length of thenucleic acids and the degree of complementation. Such variables are wellknown in the art. More specifically, the greater the degree ofsimilarity or homology between two nucleotide sequences, the greater thevalue of Tm for hybrids of nucleic acids having those sequences. Forhybrids of greater than 100 nucleotides in length, equations forcalculating Tm have been derived (see Sambrook et al., supra). Forhybridization with shorter nucleic acids, the position of mismatchesbecomes more important, and the length of the oligonucleotide determinesits specificity (see Sambrook et al., supra).

[0067] Production of the Human Δ5-Desaturase Enzyme

[0068] Once the gene encoding the human Δ5-desaturase enzyme has beenisolated, it may then be introduced into either a prokaryotic oreukaryotic host cell through the use of a vector or construct.

[0069] The vector, for example, a bacteriophage, cosmid or plasmid, maycomprise the nucleotide sequence encoding the human Δ5-desaturase enzymeas well as any promoter which is functional in the host cell and is ableto elicit expression of the human Δ5-desaturase encoded by thenucleotide sequence. The promoter is in operable association with oroperably linked to the nucleotide sequence. (A promoter is said to be“operably linked” with a coding sequence if the promoter affectstranscription or expression of the coding sequence.) Suitable promotersinclude, for example, those from genes encoding alcohol dehydrogenase,glyceraldehyde-3-phosphate dehydrogenase, phosphoglucoisomerase,phosphoglycerate kinase, acid phosphatase, T7, TPI, lactase,metallothionein, cytomegalovirus immediate early, whey acidic protein,glucoamylase, and promoters activated in the presence of galactose, forexample, GAL1 and GAL10. Additionally, nucleotide sequences which encodeother proteins, oligosaccharides, lipids, etc. may also be includedwithin the vector as well as other regulatory sequences such as apolyadenylation signal (e.g., the poly-A signal of SV-40T-antigen,ovalalbumin or bovine growth hormone). The choice of sequences presentin the construct is dependent upon the desired expression products aswell as the nature of the host cell.

[0070] As noted above, once the vector has been constructed, it may thenbe introduced into the host cell of choice by methods known to those ofordinary skill in the art including, for example, transfection,transformation and electroporation (see Molecular Cloning: A LaboratoryManual, 2^(nd) ed., Vol. 1-3, ed. Sambrook et al., Cold Spring HarborLaboratory Press (1989)). The host cell is then cultured under suitableconditions permitting expression of the desired PUFA which is thenrecovered and purified.

[0071] Examples of suitable prokaryotic host cells include, for example,bacteria such as Escherichia coli, Bacillus subtilis as well ascyanobacteria such as Spirulina spp. (i.e., blue-green algae). Examplesof suitable eukaryotic host cells include, for example, mammalian cells,plant cells, yeast cells such as Saccharomyces cerevisiae, Saccharomycescarlsbergensis, Lipomyces starkey, Candida spp. such as Yarrowia(Candida) lipolytica, Kluyveromyces spp., Pichia spp., Trichoderma spp.or Hansenula spp., or fungal cells such as filamentous fungal cells, forexample, Aspergillus, Neurospora and Penicillium. Preferably,Saccharomyces cerevisiae (baker's yeast) cells are utilized.

[0072] Expression in a host cell can be accomplished in a transient orstable fashion. Transient expression can occur from introducedconstructs which contain expression signals functional in the host cell,but which constructs do not replicate and rarely integrate in the hostcell, or where the host cell is not proliferating. Transient expressionalso can be accomplished by inducing the activity of a regulatablepromoter operably linked to the gene of interest, although suchinducible systems frequently exhibit a low basal level of expression.Stable expression can be achieved by introduction of a construct thatcan integrate into the host genome or that autonomously replicates inthe host cell. Stable expression of the gene of interest can be selectedfor through the use of a selectable marker located on or transfectedwith the expression construct, followed by selection for cellsexpressing the marker. When stable expression results from integration,the site of the construct's integration can occur randomly within thehost genome or can be targeted through the use of constructs containingregions of homology with the host genome sufficient to targetrecombination with the host locus. Where constructs are targeted to anendogenous locus, all or some of the transcriptional and translationalregulatory regions can be provided by the endogenous locus.

[0073] A transgenic mammal may also be used in order to express theenzyme of interest (i.e., the human Δ5-desaturase), and ultimately thePUFA(s) of interest. More specifically, once the above-describedconstruct is created, it may be inserted into the pronucleus of anembryo. The embryo may then be implanted into a recipient female.Alternatively, a nuclear transfer method could also be utilized(Schnieke et al., Science 278:2130-2133 (1997)). Gestation and birth arethen permitted (see, e.g., U.S. Pat. No. 5,750,176 and U.S. Patent No.5,700,671). Milk, tissue or other fluid samples from the offspringshould then contain altered levels of PUFAs, as compared to the levelsnormally found in the non-transgenic animal. Subsequent generations maybe monitored for production of the altered or enhanced levels of PUFAsand thus incorporation of the gene encoding the human Δ5-desaturaseenzyme into their genomes. The mammal utilized as the host may beselected from the group consisting of, for example, a mouse, a rat, arabbit, a pig, a goat, a sheep, a horse and a cow. However, any mammalmay be used provided it has the ability to incorporate DNA encoding theenzyme of interest into its genome.

[0074] For expression of a human Δ5-desaturase polypeptide, functionaltranscriptional and translational initiation and termination regions areoperably linked to the DNA encoding the desaturase polypeptide.Transcriptional and translational initiation and termination regions arederived from a variety of nonexclusive sources, including the DNA to beexpressed, genes known or suspected to be capable of expression in thedesired system, expression vectors, chemical synthesis, or from anendogenous locus in a host cell. Expression in a plant tissue and/orplant part presents certain efficiencies, particularly where the tissueor part is one which is harvested early, such as seed, leaves, fruits,flowers, roots, etc. Expression can be targeted to that location withthe plant by utilizing specific regulatory sequence such as those ofU.S. Pat. Nos. 5,463,174, 4,943,674, 5,106,739, 5,175,095, 5,420,034,5,188,958, and 5,589,379. Alternatively, the expressed protein can be anenzyme which produces a product which may be incorporated, eitherdirectly or upon further modifications, into a fluid fraction from thehost plant. Expression of a human Δ5-desaturase gene, or antisense humanΔ5-desaturase transcripts, can alter the levels of specific PUFAs, orderivatives thereof, found in plant parts and/or plant tissues. Thehuman Δ5-desaturase polypeptide coding region may be expressed either byitself or with other genes, in order to produce tissues and/or plantparts containing higher proportions of desired PUFAs or in which thePUFA composition more closely resembles that of human breast milk(Prieto et al., PCT publication WO 95/24494). The termination region maybe derived from the 3′ region of the gene from which the initiationregion was obtained or from a different gene. A large number oftermination regions are known to and have been found to be satisfactoryin a variety of hosts from the same and different genera and species.The termination region usually is selected as a matter of conveniencerather than because of any particular property.

[0075] As noted above, a plant (e.g., Glycine max (soybean) or Brassicanapus (canola)) or plant tissue may also be utilized as a host or hostcell, respectively, for expression of the human Δ5-desaturase enzymewhich may, in turn, be utilized in the production of polyunsaturatedfatty acids. More specifically, desired PUFAS can be expressed in seed.Methods of isolating seed oils are known in the art. Thus, in additionto providing a source for PUFAs, seed oil components may be manipulatedthrough the expression of the human Δ5-desaturase gene, as well asperhaps other desaturase genes and elongase genes, in order to provideseed oils that can be added to nutritional compositions, pharmaceuticalcompositions, animal feeds and cosmetics. Once again, a vector whichcomprises a DNA sequence encoding the human Δ5-desaturase operablylinked to a promoter, will be introduced into the plant tissue or plantfor a time and under conditions sufficient for expression of the humanΔ5-desaturase gene. The vector may also comprise one or more genes thatencode other enzymes, for example, Δ4-desaturase, elongase,Δ6-desaturase, Δ12-desaturase, Δ15-desaturase, Δ17-desaturase, and/orΔ19-desaturase. The plant tissue or plant may produce the relevantsubstrate (e.g., DGLA, GLA, EPA, 20:4n−3, etc.) upon which the enzymesact or a vector encoding enzymes which produce such substrates may beintroduced into the plant tissue, plant cell or plant. In addition,substrate may be sprayed on plant tissues expressing the appropriateenzymes. Using these various techniques, one may produce PUFAs (e.g.,n−6 unsaturated fatty acids such as AA, or n−3 fatty acids such as EPAor DHA) by use of a plant cell, plant tissue or plant. It should also benoted that the invention also encompasses a transgenic plant comprisingthe above-described vector, wherein expression of the nucleotidesequence of the vector results in production of a polyunsaturated fattyacid in, for example, the seeds of the transgenic plant.

[0076] The substrates which may be produced by the host cell eithernaturally or transgenically, as well as the enzymes which may be encodedby DNA sequences present in the vector which is subsequently introducedinto the host cell, are shown in FIG. 11.

[0077] In view of the above, the present invention encompasses a methodof producing the human Δ5-desaturase enzyme comprising the steps of: 1)isolating the nucleotide sequence of the gene encoding humanΔ5-desaturase enzyme; 2) constructing a vector comprising saidnucleotide sequence; and 3) introducing said vector into a host cellunder time and conditions sufficient for the production of thedesaturase enzyme.

[0078] The present invention also encompasses a method of producingpolyunsaturated fatty acids comprising exposing an acid to the humanΔ5-desaturase enzyme such that the desaturase converts the acid to apolyunsaturated fatty acid. For example, when 20:3n−6 is exposed tohuman Δ5-desaturase enzyme, it is converted to AA. AA may then beexposed to elongase which elongates the AA to adrenic acid (i.e.,22:4n−6). Alternatively, human Δ5-desaturase may be utilized to convert20:4n−3 to 20:5n−3 which may be exposed to elongase and converted to(n−3)-docosapentaenoic acid. The (n−3)-docosapentaenoic acid may then beconverted to DHA by use of Δ4-desaturase. Thus, human Δ5-desaturase maybe used in the production of polyunsaturated fatty acids which may beused, in turn, for particular beneficial purposes.

[0079] Uses of the Human Δ5-Desaturase Gene and Enzyme Encoded Thereby

[0080] As noted above, the isolated human Δ5-desaturase gene and thedesaturase enzyme encoded thereby have many uses. For example, the geneand corresponding enzyme may be used indirectly or directly in theproduction of polyunsaturated fatty acids, for example, AA, adrenic acidor EPA. (“Directly” is meant to encompass the situation where the enzymedirectly converts the acid to another acid, the latter of which isutilized in a composition (e.g., the conversion of DGLA to AA).“Indirectly” is meant to encompass the situation where an acid isconverted to another acid (i.e., a pathway intermediate) by thedesaturase (e.g., DGLA to AA) and then the latter acid is converted toanother acid by use of a non-desaturase enzyme (e.g., AA to adrenic acidby elongase or by use of another desaturase enzyme (e.g., AA to EPA byΔ17-desaturase.)). These polyunsaturated fatty acids (i.e., thoseproduced either directly or indirectly by activity of the desaturaseenzyme) may be added to, for example, nutritional compositions,pharmaceutical compositions, cosmetics, and animal feeds, all of whichare encompassed by the present invention. These uses are described, indetail, below.

[0081] Nutritional Compositions

[0082] The present invention includes nutritional compositions. Suchcompositions, for purposes of the present invention, include any food orpreparation for human consumption including for enteral or parenteralconsumption, which when taken into the body (a) serve to nourish orbuild up tissues or supply energy and/or (b) maintain, restore orsupport adequate nutritional status or metabolic function.

[0083] The nutritional composition of the present invention comprises atleast one oil or acid produced directly or indirectly by use of thehuman Δ5-desaturase gene, in accordance with the present invention, andmay either be in a solid or liquid form. Additionally, the compositionmay include edible macronutrients, vitamins and minerals in amountsdesired for a particular use. The amount of such ingredients will varydepending on whether the composition is intended for use with normal,healthy infants, children or adults having specialized needs such asthose which accompany certain metabolic conditions (e.g., metabolicdisorders).

[0084] Examples of macronutrients which may be added to the compositioninclude but are not limited to edible fats, carbohydrates and proteins.Examples of such edible fats include but are not limited to coconut oil,soy oil, and mono- and diglycerides. Examples of such carbohydratesinclude but are not limited to glucose, edible lactose and hydrolyzedsearch. Additionally, examples of proteins which may be utilized in thenutritional composition of the invention include but are not limited tosoy proteins, electrodialysed whey, electrodialysed skim milk, milkwhey, or the hydrolysates of these proteins.

[0085] With respect to vitamins and minerals, the following may be addedto the nutritional compositions of the present invention: calcium,phosphorus, potassium, sodium, chloride, magnesium, manganese, iron,copper, zinc, selenium, iodine, and Vitamins A, E, D, C, and the Bcomplex. Other such vitamins and minerals may also be added.

[0086] The components utilized in the nutritional compositions of thepresent invention will be of semi-purified or purified origin. Bysemi-purified or purified is meant a material which has been prepared bypurification of a natural material or by synthesis.

[0087] Examples of nutritional compositions of the present inventioninclude but are not limited to infant formulas, dietary supplements,dietary substitutes, and rehydration compositions. Nutritionalcompositions of particular interest include but are not limited to thoseutilized for enteral and parenteral supplementation for infants,specialist infant formulas, supplements for the elderly, and supplementsfor those with gastrointestinal difficulties and/or malabsorption.

[0088] The nutritional composition of the present invention may also beadded to food even when supplementation of the diet is not required. Forexample, the composition may be added to food of any type including butnot limited to margarines, modified butters, cheeses, milk, yogurt,chocolate, candy, snacks, salad oils, cooking oils, cooking fats, meats,fish and beverages.

[0089] In a preferred embodiment of the present invention, thenutritional composition is an enteral nutritional product, morepreferably, an adult or pediatric enteral nutritional product. Thiscomposition may be administered to adults or children experiencingstress or having specialized needs due to chronic or acute diseasestates. The composition may comprise, in addition to polyunsaturatedfatty acids produced in accordance with the present invention,macronutrients, vitamins and minerals as described above. Themacronutrients may be present in amounts equivalent to those present inhuman milk or on an energy basis, i.e., on a per calorie basis.

[0090] Methods for formulating liquid or solid enteral and parenteralnutritional formulas are well known in the art. (See also the Examplesbelow.)

[0091] The enteral formula, for example, may be sterilized andsubsequently utilized on a ready-to-feed (RTF) basis or stored in aconcentrated liquid or powder. The powder can be prepared by spraydrying the formula prepared as indicated above, and reconstituting it byrehydrating the concentrate. Adult and pediatric nutritional formulasare well known in the art and are commercially available (e.g.,Similac®, Ensure®, Jevity® and Alimentum® from Ross Products Division,Abbott Laboratories, Columbus, Ohio). An oil or acid produced inaccordance with the present invention may be added to any of theseformulas.

[0092] The energy density of the nutritional compositions of the presentinvention, when in liquid form, may range from about 0.6 Kcal to about 3Kcal per ml. When in solid or powdered form, the nutritional supplementsmay contain from about 1.2 to more than 9 Kcals per gram, preferablyabout 3 to 7 Kcals per gm. In general, the osmolality of a liquidproduct should be less than 700 mOsm and, more preferably, less than 660mOsm.

[0093] The nutritional formula may include macronutrients, vitamins, andminerals, as noted above, in addition to the PUFAs produced inaccordance with the present invention. The presence of these additionalcomponents helps the individual ingest the minimum daily requirements ofthese elements. In addition to the provision of PUFAs, it may also bedesirable to add zinc, copper, folic acid and antioxidants to thecomposition. It is believed that these substance boost a stressed immunesystem and will therefore provide further benefits to the individualreceiving the composition. A pharmaceutical composition may also besupplemented with these elements.

[0094] In a more preferred embodiment, the nutritional compositioncomprises, in addition to antioxidants and at least one PUFA, a sourceof carbohydrate wherein at least 5 weight percent of the carbohydrate isindigestible oligosaccharide. In a more preferred embodiment, thenutritional composition additionally comprises protein, taurine, andcarnitine.

[0095] As noted above, the PUFAs produced in accordance with the presentinvention, or derivatives thereof, may be added to a dietary substituteor supplement, particularly an infant formula, for patients undergoingintravenous feeding or for preventing or treating malnutrition or otherconditions or disease states. As background, it should be noted thathuman breast milk has a fatty acid profile comprising from about 0.15%to about 0.36% as DHA, from about 0.03% to about 0.13% as EPA, fromabout 0.30% to about 0.88% as AA, from about 0.22% to about 0.67% asDGLA, and from about 0.27% to about 1.04% as GLA. Thus, fatty acids suchas AA, EPA and/or docosahexaenoic acid (DHA), produced in accordancewith the present invention, can be used to alter, for example, thecomposition of infant formulas in order to better replicate the PUFAcontent of human breast milk or to alter the presence of PUFAs normallyfound in a non-human mammal's milk. In particular, a composition for usein a pharmacologic or food supplement, particularly a breast milksubstitute or supplement, will preferably comprise one or more of AA,DGLA and GLA. More preferably, the oil will comprise from about 0.3 to30% AA, from about 0.2 to 30% DGLA, and/or from about 0.2 to about 30%GLA.

[0096] Parenteral nutritional compositions comprising from about 2 toabout 30 weight percent fatty acids calculated as triglycerides areencompassed by the present invention. The preferred composition hasabout 1 to about 25 weight percent of the total PUFA composition as GLA(U.S. Pat. No. 5,196,198). Other vitamins, particularly fat-solublevitamins such as vitamin A, D, E and L-carnitine can optionally beincluded. When desired, a preservative such as alpha-tocopherol may beadded in an amount of about 0.1% by weight.

[0097] In addition, the ratios of AA, DGLA and GLA can be adapted for aparticular given end use. When formulated as a breast milk supplement orsubstitute, a composition which comprises one or more of AA, DGLA andGLA will be provided in a ratio of about 1:19:30 to about 6:1:0.2,respectively. For example, the breast milk of animals can vary in ratiosof AA:DGLA:GLA ranging from 1:19:30 to 6:1:0.2, which includesintermediate ratios which are preferably about 1:1:1, 1:2:1, 1:1:4. Whenproduced together in a host cell, adjusting the rate and percent ofconversion of a precursor substrate such as GLA and DGLA to AA can beused to precisely control the PUFA ratios. For example, a 5% to 10%conversion rate of DGLA to AA can be used to produce an AA to DGLA ratioof about 1:19, whereas a conversion rate of about 75% TO 80% can be usedto produce an AA to DGLA ratio of about 6:1. Therefore, whether in acell culture system or in a host animal, regulating the timing, extentand specificity of human Δ5-desaturase expression, as well as theexpression of other desaturases and elongases, can be used to modulatePUFA levels and ratios. The PUFAs/acids produced in accordance with thepresent invention (e.g., AA and EPA) may then be combined with otherPUFAs/acids (e.g., GLA) in the desired concentrations and ratios.

[0098] Additionally, PUFA produced in accordance with the presentinvention or host cells containing them may also be used as animal foodsupplements to alter an animal's tissue or milk fatty acid compositionto one more desirable for human or animal consumption.

[0099] Pharmaceutical Compositions

[0100] The present invention also encompasses a pharmaceuticalcomposition comprising one or more of the acids and/or resulting oilsproduced using the human Δ5-desaturase gene, in accordance with themethods described herein. More specifically, such a pharmaceuticalcomposition may comprise one or more of the acids and/or oils as well asa standard, well-known, non-toxic pharmaceutically acceptable carrier,adjuvant or vehicle such as, for example, phosphate buffered saline,water, ethanol, polyols, vegetable oils, a wetting agent or an emulsionsuch as a water/oil emulsion. The composition may be in either a liquidor solid form. For example, the composition may be in the form of atablet, capsule, ingestible liquid or powder, injectible, or topicalointment or cream. Proper fluidity can be maintained, for example, bythe maintenance of the required particle size in the case of dispersionsand by the use of surfactants. It may also be desirable to includeisotonic agents, for example, sugars, sodium chloride and the like.Besides such inert diluents, the composition can also include adjuvants,such as wetting agents, emulsifying and suspending agents, sweeteningagents, flavoring agents and perfuming agents.

[0101] Suspensions, in addition to the active compounds, may comprisesuspending agents such as, for example, ethoxylated isostearyl alcohols,polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanthor mixtures of these substances.

[0102] Solid dosage forms such as tablets and capsules can be preparedusing techniques well known in the art. For example, PUFAs produced inaccordance with the present invention can be tableted with conventionaltablet bases such as lactose, sucrose, and cornstarch in combinationwith binders such as acacia, cornstarch or gelatin, disintegratingagents such as potato starch or alginic acid, and a lubricant such asstearic acid or magnesium stearate. Capsules can be prepared byincorporating these excipients into a gelatin capsule along withantioxidants and the relevant PUFA(s). The antioxidant and PUFAcomponents should fit within the guidelines presented above.

[0103] For intravenous administration, the PUFAs produced in accordancewith the present invention or derivatives thereof may be incorporatedinto commercial formulations such as Intralipids™. The typical normaladult plasma fatty acid profile comprises 6.64 to 9.46% of AA, 1.45 to3.11% of DGLA, and 0.02 to 0.08% of GLA. These PUFAs or their metabolicprecursors can be administered alone or in combination with other PUFAsin order to achieve a normal fatty acid profile in a patient. Wheredesired, the individual components of the formulations may be providedindividually, in kit form, for single or multiple use. A typical dosageof a particular fatty acid is from 0.1 mg to 20 g (up to 100 g) dailyand is preferably from 10 mg to 1, 2, 5 or 10 g daily.

[0104] Possible routes of administration of the pharmaceuticalcompositions of the present invention include, for example, enteral(e.g., oral and rectal) and parenteral. For example, a liquidpreparation may be administered, for example, orally or rectally.Additionally, a homogenous mixture can be completely dispersed in water,admixed under sterile conditions with physiologically acceptablediluents, preservatives, buffers or propellants in order to form a sprayor inhalant.

[0105] The route of administration will, of course, depend upon thedesired effect. For example, if the composition is being utilized totreat rough, dry, or aging skin, to treat injured or burned skin, or totreat skin or hair affected by a disease or condition, it may perhaps beapplied topically.

[0106] The dosage of the composition to be administered to the patientmay be determined by one of ordinary skill in the art and depends uponvarious factors such as weight of the patient, age of the patient,immune status of the patient, etc.

[0107] With respect to form, the composition may be, for example, asolution, a dispersion, a suspension, an emulsion or a sterile powderwhich is then reconstituted.

[0108] The present invention also includes the treatment of variousdisorders by use of the pharmaceutical and/or nutritional compositionsdescribed herein. In particular, the compositions of the presentinvention may be used to treat restenosis after angioplasty.Furthermore, symptoms of inflammation, rheumatoid arthritis, asthma andpsoriasis may also be treated with the compositions of the invention.Evidence also indicates that PUFAs may be involved in calciummetabolism; thus, the compositions of the present invention may,perhaps, be utilized in the treatment or prevention of osteoporosis andof kidney or urinary tract stones.

[0109] Additionally, the compositions of the present invention may alsobe used in the treatment of cancer. Malignant cells have been shown tohave altered fatty acid compositions. Addition of fatty acids has beenshown to slow their growth, cause cell death and increase theirsusceptibility to chemotherapeutic agents. Moreover, the compositions ofthe present invention may also be useful for treating cachexiaassociated with cancer.

[0110] The compositions of the present invention may also be used totreat diabetes (see U.S. Pat. No. 4,826,877 and Horrobin et al., Am. J.Clin. Nutr. Vol. 57 (Suppl.) 732S-737S). Altered fatty acid metabolismand composition have been demonstrated in diabetic animals.

[0111] Furthermore, the compositions of the present invention,comprising PUFAs produced either directly or indirectly through the useof the human Δ5-desaturase enzyme, may also be used in the treatment ofeczema, in the reduction of blood pressure, and in the improvement ofmathematics examination scores. Additionally, the compositions of thepresent invention may be used in inhibition of platelet aggregation,induction of vasodilation, reduction in cholesterol levels, inhibitionof proliferation of vessel wall smooth muscle and fibrous tissue(Brenner et al., Adv. Exp. Med. Biol. Vol. 83, p.85-101, 1976),reduction or prevention of gastrointestinal bleeding and other sideeffects of non-steroidal anti-inflammatory drugs (see U.S. Pat. No.4,666,701), prevention or treatment of endometriosis and premenstrualsyndrome (see U.S. Pat. No. 4,758,592), and treatment of myalgicencephalomyelitis and chronic fatigue after viral infections (see U.S.Pat. No. 5,116,871).

[0112] Further uses of the compositions of the present invention includeuse in the treatment of AIDS, multiple sclerosis, and inflammatory skindisorders, as well as for maintenance of general health.

[0113] Additionally, the composition of the present invention may beutilized for cosmetic purposes. It may be added to pre-existing cosmeticcompositions such that a mixture is formed or may be used as a solecomposition.

[0114] Veterinary Applications

[0115] It should be noted that the above-described pharmaceutical andnutritional compositions may be utilized in connection with animals(i.e., domestic or non-domestic), as well as humans, as animalsexperience many of the same needs and conditions as humans. For example,the oil or acids of the present invention may be utilized in animal feedsupplements, animal feed substitutes, animal vitamins or in animaltopical ointments.

[0116] The present invention may be illustrated by the use of thefollowing non-limiting examples:

EXAMPLE I Human Desaturase Gene Sequences

[0117] As described in International Application PCT/US98/07422 (hereinincorporated in its entirety by reference), the putative humandesaturase gene sequences involved in long chain polyunsaturated fattyacid biosynthesis were isolated based on homology between the human cDNAsequences and Mortierella alpina desaturase gene sequences. The threeconserved “histidine boxes” known to be conserved among membrane-bounddesaturases were found. As with other membrane-bound desaturases, thefinal HXXHH histidine box motif was found to be QXXHH. The amino acidsequence of the putative human desaturases exhibited homology to M.alpina Δ5-, Δ6-, Δ9-, and Δ12-desaturases.

[0118] The M. alpina Δ5-desaturase and Δ6-desaturase cDNA sequences wereused to search the LifeSeq database of Incyte Pharmaceuticals, Inc.,Palo Alto, Calif. The Δ5-desaturase sequence was divided intofragments: 1) amino acid no. 1-150, 2) amino acid no. 151-300, and 3)amino acid no. 301-446. The Δ6 desaturase sequence was divided intothree fragments: 1) amino acid no. 1-150, 2) amino acid no. 151-300, and3) amino acid no. 301-457. These polypeptide fragments were searchedagainst the database using the “tblastn” algorithm. This algorithmcompares a protein query sequence against a nucleotide sequence databasedynamically translated in all six reading frames (both strands).

[0119] The polypeptide fragments 2 and 3 of M. alpina Δ5- andΔ6-desaturases have homologies with the CloneID sequences as outlined inFIG. 1. The CloneID represents an individual sequence from the IncyteLifeSeq database. After the “tblastn” results had been reviewed, CloneInformation was searched with the default settings of Stringencyof >=50, and Productscore <=100 for different CloneID numbers. The CloneInformation Results displayed the information including the ClusterID,CloneID, Library, HitID, and Hit Description. When selected, theClusterID number displayed the clone information of all the clones thatbelong in that ClusterID. The Assemble command assembled all of theCloneID which comprise the ClusterID. The following default setting wereused for GCG (Genetics Computer Group, University of WisconsinBiotechnology Center, Madison, Wis.) Assembly:

[0120] Word Size: 7; Minimum Overlap: 14; Stringency: 0.8;

[0121] Minimum Identity: 14; Maximum Gap: 10; Gap Weight: 8; and

[0122] Length Weight: 2.

[0123] GCG Assembly Results displayed the contigs generated on the basisof sequence information within the CloneID. A contig is an alignment ofDNA sequences based on areas of homology among these sequences. A newsequence (consensus sequence) was generated based on the aligned DNAsequence within a contig. The contig. containing the CloneID wasidentified, and the ambiguous sites of the consensus sequence wereedited based on the alignment of the CloneIDs (see FIGS. 2-6) togenerate the best possible sequence. The procedure was repeated for allsix CloneID listed in FIG. 1. This produced five unique contigs. Theedited consensus sequences of the 5 contigs were imported into theSequencher software program (Gene Codes Corporation, Ann Arbor, Mich.).These consensus sequences were assembled. The contig 2511785 overlapswith contig 3506132, and this new contig was called 2535 (FIG. 7). Thecontigs from the Sequencher program were copied into the SequenceAnalysis software package of GCG.

[0124] Each contig was translated in all six reading frames into proteinsequences. The M. alpina Δ5-desaturase (Ma29) and Δ6-desaturase (Ma524)sequences were compared with each of the translated contigs using theFastA search (a Pearson and Lipman search for similarity between a querysequence and a group of sequences of the same type (nucleic acid orprotein)). Homology among these sequences suggest the open readingframes of each contig as underlined in FIGS. 3, 5, and 7. The homologyamong the M. alpina Δ5- and Δ6-desaturase sequences to contigs 2535 and3854933 were utilized to create the final contig called 253538a (seeFIG. 8). FIG. 9 is the FastA match of the translated sequences of thefinal contig 253538a and Ma29, and FIG. 10 is the FastA match of thetranslated sequences of the final contig 253538a and Ma524.

[0125] Although the open reading frame was generated by merging the twocontigs, the contig 2535 shows that there is a unique sequence in thebeginning of this contig which does not match with the contig 3854933.Therefore, it is possible that these contigs were generated fromindependent desaturase-like human genes.

[0126] The contig 253538a contains an open reading frame encoding 432amino acid (FIG. 8, underlined). It starts with Gln (CAG) and ends withthe stop codon (TGA) (both in bold). The contig 253538a aligns with bothM. alpina Δ5- and Δ6-desaturase sequences, suggesting that it could beeither of the desaturases, as well as other known desaturases whichshare homology with each other. The individual contigs listed in FIG. 1,as well as the intermediate contig 2535 and the final contig 253538a canbe utilized to isolate the complete genes for human desaturases.

[0127] Determination of Human Δ5-Desaturase Gene Sequence

[0128] Primers RO384 and RO388 were designed based on the 5′ and 3′sequences, respectively, of contig 2535. The human monocyte cDNA library(Clontech, Palo Alto, Calif.) was amplified with the vector primer RO329(5′-CAG ACC AAC TGG TAA TGG TAG-3′) and RO384 (5′-TCA GGC CCA AGC TGGATG GCT GCA ACA TG-3′), and also with the vector primer RO328 (5′-CTCCTG GAG CCC GTC AGT ATC-3′) and RO388 (5′-ATG GTG GGG AAG AGG TGG TGCTCA ATC TG-3′). Polymerase Chain Reaction (PCR) was carried out in a 100μl volume containing: 1 μl of human monocyte cDNA library, 10 pM eachprimer, 10 μl of 10× buffer and 1.0 U of Taq Polymerase. Thermocyclerconditions in Perkin Elmer 9600 were as follows:

[0129] 94° C. for 2 mins, then 30 cycles of 94° C. for 1 min., 58° C.for 2 mins. and 72° C. for 3 mins. PCR was followed by an additionalextension at 72° C. for 7 minutes.

[0130] The PCR amplified mixture was run on a gel, and the amplifiedfragments were gel purified. The isolated fragment from PCRamplification with RO329 and RO384 was approximately 900 bp, and thatfrom PCR amplification with RO328 and RO388 was approximately 650 bp.These isolated fragments were filled-in using T4 DNA polymerase, and thefilled-in fragments were cloned into the PCR-Blunt vector (InvitrogenCorp., Carlsbad, Calif.). The clone of RO329/RO384 amplified fragmentwas designated as pRAE-7, and the clone of RO328/RO388 amplifiedfragment was designated as pRAE-8. Both ends of the clones weresequenced using ABI 373 DNA Sequencer (Applied Biosystems, Foster City,Calif.) and assembled using the Sequencher program (a sequence analysisprogram, Gene Codes Corporation, Ann Arbor, Mich.). This assembly of thesequences revealed that the two clones contained different sizes of thesame gene (FIG. 14). The complete sequence of the pRAE-7 gene wascompiled (FIG. 15) and searched against the known sequences in thepublic database.

[0131] The FastA algorithm is a Pearson and Lipman search for similaritybetween a query sequence and a group of sequences of the same type(nucleic acid or protein). The pRAE-7 gene sequence was translated insix reading frames, and using this method, the Swissprot database(Genetics Computer Group (GCG) (Madison, Wis.) was searched. The gene inpRAE-7 was identified as a putative human desaturase based on itshomology to known desaturases. The Swissprot database search producedmatches against the omega-3 fatty acid desaturase from mung bean (23.4%identity in 303 AA overlap), linoleoyl-CoA desaturase from Synechocystissp. (24.3% identity in 280 AA overlap), omega-6 fatty acid desaturasefrom soybean (19.7% identity in 284 AA overlap), and acyl-CoA desaturase1 from Saccharomyces cerevisiae (21.6% identity in 134 AA overlap). TheFastA search against the M. alpina desaturases produced matches againstthe Δ6-(31.9% identity in 285 AA overlap), the Δ5-(28.4% identity in 292AA overlap), and the Δ12-(23.0% identity in 274 AA overlap) desaturases.The matched sequence alignment of the putative human desaturase gene inpRAE-7 against M. alpina Δ5-desaturase (Ma29), M. alpina Δ6-desaturase(Ma524) as well as to the contigs 2535 and 38 are displayed in FIGS. 16,17, 18, and 19 respectively.

[0132] The contigs 2535, 38, and 253538a were generated based onassemblies of various sequences as well as their homologies against theknown desaturases. However, upon examining FIGS. 18 and 19, it can beconcluded that the contigs are merely indications as to what thesequences of the human desaturases might possibly be.

[0133] The 5′ end of the gene, the ATG (Methionine), is necessary forexpressing the human desaturase in yeast. FIGS. 16 and 17 show thatpRAE-7 is probably just the last ⅔ of a desaturase gene. Several of theomega-3 and omega-6 fatty acid desaturases, as well as the linoleoyl-CoAdesaturase mentioned above, are smaller than the M. alpina Δ5- andΔ6-desaturases, ranging in sizes of 359-380 amino acids. It wasconcluded from all of the sequences evaluated thus far that the isolatedgene probably needed anywhere from 180-480 bp (60-160 amino acids) ofadditional 5′ sequence for expressing a complete enzyme.

[0134] In order to extend the 5′ sequence of the human desaturase gene,the Marathon cDNA Amplification Kit (Clontech, Palo Alto, Calif.) wasused to screen the human liver marathon ready cDNA (Clontech). The rapidamplification of cDNA ends (RACE) reactions are efficient for both 5′and 3′ long-distance PCR. Following the 5′ RACE protocol outlined in thekit, the primers RO430 (5′-GTG GCT GTT GTT ATT GGT GAA GAT AGG CAT C-3′)(designed based on the pRAE-7 gene 3′ sequence, downstream of the TAA(stop)) and the marathon adaptor primer (AP1) from the kit, were used togenerate three PCR amplified products, which were designated A, B, andC. The fragment sizes were approximately 1.5 Kb, 1.4 Kb, 1.2 Kb,respectively. The fragments were filled-in with T4 DNA polymerase, andcloned into the pCR-blunt vector. A total of twenty-two clones weregenerated and sequenced. Using the FastA algorithm, the sequences weresearched against the GenEMBL database of GCG.

[0135] Many of the sequences had a great homology to the human DNAsequence with the GenBank accession number of AC004228. This DNAsequence is described as: Sequencing in Progress, Homo sapiensChromosome 11q12pac pDJ519o3; HTGS phase 1, 18 unordered pieces. The 18contigs were recorded in an arbitrary fashion. Using this sequenceinformation and the information from the assembled sequences of theclones, the clones were categorized into five groups.

[0136] All of the clones have the same sequence downstream of the BamHIsite (see FIG. 12, underlined). But each group represents a different 5′sequence, with a total of 10 clones being too short to be the fulllength gene. Group 1, represented by clone A-1, is comprised of 5 cloneswhich have homology to cytochrome b5 gene (FIG. 20). A translationalstart codon, ATG, is not present in clone A-1; however, as can be seenin FIG. 21, there is an ATG (underlined) present in the ac004228sequence 17 bp upstream of the strong area of homology between A-1 andac004228. Starting from the strong area of homology, A-1 has an openreading frame of 1318 bp. However, starting from the ATG, the openreading frame is 1335 bp. Group 2, represented by clone 3-5, iscomprised of 3 clones which have an ATG within an NcoI site, but fourtranslational stop codons between the ATG and the BamHI site (FIG. 22,the NcoI, BamHI sites are in bold, and the four termination codons areunderlined). Group 3 is comprised of one clone, A-10, which has an ATG135 bp upstream of the BamHI site, giving an open reading frame of 1267bp (FIG. 23). Group 4 is comprised of 2 clones, represented by cloneA-16, which does not have an ATG; however, upstream of where thesequence aligns with ac004228, there is an ATG (FIG. 24, underlined).The open reading frame of this group is 1347 bp. Group 5 is comprised ofone clone which does not have an ATG. However, this clone matches theac004228 sequence even upstream of the BamHI site (FIG. 25).

[0137] As illustrated in FIG. 26, many of the clones from the fivegroups are represented in order with the ac004228 sequence. Thereappeared to be a high level of splicing, with the sequence downstream ofthe BamHI site (in bold) acting as the common anchor for the various 5′exons. All of the potential start sites are also in bold, and thesequences found within the clones have been underlined.

[0138] The A-1 sequence was used to search the LifeSeq database ofIncyte Pharmaceuticals, Inc., Palo Alto, Calif., to see if its latestversion would also have sequences with homology to our desaturase genesequence. Two contigs were generated in this search, contig 3381584 andcontig 2153526. The human desaturase gene sequence was initiallycompiled based on sequences from Group I clones and ac004228. However,FIG. 12 represents the actual DNA sequence of the isolated gene. TheIncyte contigs were used to confirm this sequence (see FIGS. 27 and 28).The human desaturase translated sequence, consisting of 445 amino acids(FIG. 13), was also matched with the original contigs 253538a and 38.These alignments are shown in FIGS. 29 and 30, respectively.

[0139] The FastA search of the human desaturase gene against theSwissprot database produced matches against the omega-3 fatty aciddesaturase from mung bean (22.4% identity in 381 AA overlap),linoleoyl-CoA desaturase from Synechocystis Sp. (24.5% identity in 335AA overlap), omega-6 fatty acid desaturase from soybean (20.3% identityin 290 AA overlap), and acyl-CoA desaturase 1 from Saccharomycescerevisiae (21.4% identity in 168 AA overlap). The FastA search againstM. alpina desaturases produced matches against the Δ6-(30.5% identity in455 AA overlap), Δ5-(27.5% identity in 455 AA overlap), andΔ12-desaturases (22.5% identity in 382 AA overlap). The FastA match ofthe human desaturase translated sequence against the ma524 (M. alpinaΔ6-desaturase) and ma29 (M. alpina Δ5-desaturase) sequences are shown inFIGS. 31 and 32, respectively.

EXAMPLE II Construction of Clones

[0140] New clones were generated based on clones from three of theGroups mentioned above, clones A-1, A-10, and A-16. Two primers whichwere modified with 5′ phosphate, RO526 (5′-CAT GGC CCC CGA CCC GGTGG-3′) and RO527 (5′-GCG GCC ACC GGG TCG GGG GC-3′), were annealedtogether to form an adaptor. This adaptor which has NcoI and BsaIoverhangs, were ligated with the A-1 clone, which had been cut withBsaI/HindIII and gel purified, for 15 min at room temperature. ThepYX242(NcoI/HindIII) vector (Novagen, Madison, Wis.) was added to thisligation mixture and allowed to incubate at room temperature for anadditional 45 min. This produced a clone designated as pRAE-28-5.(Plasmid pRAE-28-5 was deposited with the American Type CultureCollection, 10801 University Boulevard, Manassas, Va. 20110-2209 on Dec.21, 1998, under the terms of the Budapest Treaty, and was accorded ATCCnumber ______.)

[0141] The A-10 clone was PCR amplified with RO512 (5′-GAT TGG GTG CCATGG GGA TGC GGG ATG AAA AGG C-3′) and RO5 (5′-GAA ACA GCT ATG ACCATG-3′), the amplified product was cut with NcoI and HindIII and gelpurified, and the purified fragment was cloned into pYX242(NcoI/HindIII). This new clone was designated as pRAE-26-1.

[0142] The A-10 clone was also PCR amplified with RO580 (5′-TCC TGC GAATTC ACC ATG AAA AGG CGG GAG AGA G-3′) and RO5, the amplified product wascut with NcoI and HindIII and gel purified, and the purified fragmentwas cloned into pYX242 (NcoI/HindIII). This new clone was designated aspRAE-33.

[0143] Two primers which were modified with 5′ phosphate, RO578 (5′-CATGGC TAG GAG AGG CAG CGC AGC CGC GTC TGG AC-3′) and RO579 (5′-CTA GGT CCAGAC GCG GCT GCG CTG CCT CTC CTA GC-3′), were annealed together to forman adaptor. This adaptor which has NcoI and BlnI overhangs, were ligatedwith the A-16 clone, which had been cut with BlnI/HindIII and gelpurified, for 15 min at room temperature. The pYX242(NcoI/HindIII)vector was added to this ligation mixture and allowed to incubate atroom temperature for an additional 45 min. This produced a clonedesignated as pRAE-35.

EXAMPLE III Expression of Human Δ5-Desaturase

[0144] The constructs pRAE-26-1, pPAE-28-5, pRAE-33, and pRAE-35 weretransformed into S. cerevisiae 334 and screened for desaturase activity.The substrates DGLA (20:3n−6), OA(18:1n−9), AA(20:4n−6), and LA(18:2n−6)were used to determine the activity of the expressed gene fromconstructs pRAE-26-1 and pRAE-28-5. Only the substrate DGLA was used todetermine the activity of the expressed gene from all of the constructs.The negative control strain was S. cerevisiae 334 containing theunaltered pYX242 vector. The cultures were grown for 48 hours at 30° C.,in selective media (Ausubel et al., Short Protocols in MolecularBiology, Ch. 13, P. 3-5 (1992)), in the presence of a particularsubstrate. Lipid fractions of each culture were extracted for analysis.The desaturase activity results are provided in FIGS. 33 and 34.

[0145] All of the values in FIG. 33 are the average of two separatesamples per strain, tested in the same run. The substrate, as well asthe fatty acid it was converted to, is shown in bold. The expressed genein the strain 334 (pRAE-28-5) is a AS-desaturase. It converted thesubstrate DGLA to a higher percent of AA than the control strain334(pYX242), 0.127% vs. 0.062%, respectively. The percent of AA presentin the cultures of strains 334(pRAE-26-1), 334(pRAE-33), and334(pRAE-35) are comparable to that of the control strain (0.075%,0.062%, and 0.063%, respectively). Therefore, it can be concluded thatthe cyt b5 sequence containing gene in the construct pRAE-28-5 expressesan active human Δ5-desaturase; whereas, the other variations of the genedo not.

[0146] The activity of the human Δ5-desaturase was further confirmed inthe experiment outlined in FIG. 34. Included in this figure are thefatty acid profiles of the strains 334 (pRAE-28-5), 334 (pRAE-26-1), andthe control strain 334 (pYX242) when DGLA(20:3n−6), OA(18:1n−9),AA(20:4n−6), or LA(18:2n−6) was used as the substrate, as well as whenno substrate was added. Again, the strain 334(pRAE-28-5) expressed anactive human Δ5-desaturase, converting DGLA to AA at a higher percentthan the control strain, 0.106% vs. 0.065%, respectively. The strain334(pRAE-26-1) had about the same amount of AA (0.06%) as the control.The conversion of the substrate OA to LA was not detected, confirmingthat the strains do not have a Δ12-desaturase activity. The conversionof the substrate AA to eicosapentaenoic acid (EPA, 20:5n−3) wasdetected, but at a very low level equal to that of the control strain,confirming that the strains do not have a Δ17-desaturase activity. Theconversion of the substrate LA to GLA was detected, but again at a verylow level equal to the control strain, confirming that the strains donot have a Δ6-desaturase activity.

[0147] The present sequence (FIG. 12) differs from the Genbank sequenceg3169158 of the LifeSeq database with respect to two positions. Inparticular, with respect to the nucleotide sequence of sequenceg3169158, position 1082 is an adenosine; however, in the presentsequence position 1082 is a thymine (see FIG. 12). Furthermore, position1229 of sequence g3169158 is an adenine whereas in the present sequenceposition 1229 is a guanine. In terms of an amino acid sequencecomparison, position 361 of the present sequence is a leucine (see FIG.13), and position 361 of sequence g3169158 is a glutamine. Furthermore,position 410 of the present sequence is an arginine, whereas position410 of sequence g3169158 is a histidine. Additionally, sequence g3169158is described, in the database, as a “hypothetical protein” which“exhibits similarity to motifs found in delta 6 desaturase, ahypothetical cytochrome b5 containing fusion protein.”However, asdemonstrated in the above example, the protein encoded by the sequencein FIG. 12 is a human Δ5-desaturase, not a Δ6-desaturase.

Nutritional Compositions

[0148] The PUFAs described in the Detailed Description may be utilizedin various nutritional supplements, infant formulations, nutritionalsubstitutes and other nutritional solutions.

[0149] I. Infant Formulations

[0150] A. Isomil® Soy Formula with Iron:

[0151] Usage: As a beverage for infants, children and adults with anallergy or sensitivity to cows milk. A feeding for patients withdisorders for which lactose should be avoided: lactase deficiency,lactose intolerance and galactosemia.

[0152] Features:

[0153] Soy protein isolate to avoid symptoms of cow's-milk-proteinallergy or sensitivity.

[0154] Lactose-free formulation to avoid lactose-associated diarrhea.

[0155] Low osmolality (240 mOs/kg water) to reduce risk of osmoticdiarrhea.

[0156] Dual carbohydrates (corn syrup and sucrose) designed to enhancecarbohydrate absorption and reduce the risk of exceeding the absorptivecapacity of the damaged gut.

[0157] 1.8 mg of Iron (as ferrous sulfate) per 100 Calories to helpprevent iron deficiency.

[0158] Recommended levels of vitamins and minerals.

[0159] Vegetable oils to provide recommended levels of essential fattyacids.

[0160] Milk-white color, milk-like consistency and pleasant aroma.

[0161] Ingredients: (Pareve) 85% water, 4.9% corn syrup, 2.6% sugar(sucrose), 2.1% soy oil, 1.9% soy protein isolate, 1.4% coconut oil,0.15% calcium citrate, 0. 11% calcium phosphate tribasic, potassiumcitrate, potassium phosphate monobasic, potassium chloride, mono- anddisglycerides, soy lecithin, carrageenan, ascorbic acid, L-methionine,magnesium chloride, potassium phosphate dibasic, sodium chloride,choline chloride, taurine, ferrous sulfate, m-inositol, alpha-tocopherylacetate, zinc sulfate, L-carnitine, niacinamide, calcium pantothenate,cupric sulfate, vitamin A palmitate, thiamine chloride hydrochloride,riboflavin, pyridoxine hydrochloride, folic acid, manganese sulfate,potassium iodide, phylloquinone, biotin, sodium selenite, vitamin D3 andcyanocobalamin.

[0162] B. Isomil® DF Soy Formula for Diarrhea:

[0163] Usage: As a short-term feeding for the dietary management ofdiarrhea in infants and toddlers.

[0164] Features:

[0165] First infant formula to contain added dietary fiber from soyfiber specifically for diarrhea management.

[0166] Clinically shown to reduce the duration of loose, watery stoolsduring mild to severe diarrhea in infants.

[0167] Nutritionally complete to meet the nutritional needs of theinfant.

[0168] Soy protein isolate with added L-methionine meets or exceeds aninfant's requirement for all essential amino acids.

[0169] Lactose-free formulation to avoid lactose-associated diarrhea.

[0170] Low osmolality (240 mOsm/kg water) to reduce the risk of osmoticdiarrhea.

[0171] Dual carbohydrates (corn syrup and sucrose) designed to enhancecarbohydrate absorption and reduce the risk of exceeding the absorptivecapacity of the damaged gut.

[0172] Meets or exceeds the vitamin and mineral levels recommended bythe Committee on Nutrition of the American Academy of Pediatrics andrequired by the Infant Formula Act.

[0173] 1.8 mg of iron (as ferrous sulfate) per 100 Calories to helpprevent iron deficiency.

[0174] Vegetable oils to provide recommended levels of essential fattyacids.

[0175] Ingredients: (Pareve) 86% water, 4.8% corn syrup, 2.5% sugar(sucrose), 2.1% soy oil, 2.0% soy protein isolate, 1.4% coconut oil,0.77% soy fiber, 0.12% calcium citrate, 0.11% calcium phosphatetribasic, 0.10% potassium citrate, potassium chloride, potassiumphosphate monobasic, mono and diglycerides, soy lecithin, carrageenan,magnesium chloride, ascorbic acid, L-methionine, potassium phosphatedibasic, sodium chloride, choline chloride, taurine, ferrous sulfate,m-inositol, alpha-tocopheryl acetate, zinc sulfate, L-carnitine,niacinamide, calcium pantothenate, cupric sulfate, vitamin A palmitate,thiamine chloride hydrochloride, riboflavin, pyridoxine hydrochloride,folic acid, manganese sulfate, potassium iodide, phylloquinone, biotin,sodium selenite, vitamin D3 and cyanocobalamin.

[0176] C. Isomil® SF Sucrose-Free Soy Formula with Iron:

[0177] Usage: As a beverage for infants, children and adults with anallergy or sensitivity to cow's-milk protein or an intolerance tosucrose. A feeding for patients with disorders for which lactose andsucrose should be avoided.

[0178] Features:

[0179] Soy protein isolate to avoid symptoms of cow's-milk-proteinallergy or sensitivity.

[0180] Lactose-free formulation to avoid lactose-associated diarrhea(carbohydrate source is Polycose® Glucose Polymers).

[0181] Sucrose free for the patient who cannot tolerate sucrose.

[0182] Low osmolality (180 mOsm/kg water) to reduce risk of osmoticdiarrhea.

[0183] 1.8 mg of iron (as ferrous sulfate) per 100 Calories to helpprevent iron deficiency.

[0184] Recommended levels of vitamins and minerals.

[0185] Vegetable oils to provide recommended levels of essential fattyacids.

[0186] Milk-white color, milk-like consistency and pleasant aroma.

[0187] Ingredients: (Pareve) 75% water, 11.8% hydrolized cornstarch,4.1% soy oil, 4.1% soy protein isolate, 2.8% coconut oil, 1.0% modifiedcornstarch, 0.38% calcium phosphate tribasic, 0. 17% potassium citrate,0.13% potassium chloride, mono- and diglycerides, soy lecithin,magnesium chloride, abscorbic acid, L-methionine, calcium carbonate,sodium chloride, choline chloride, carrageenan, taurine, ferroussulfate, m-inositol, alpha-tocopheryl acetate, zinc sulfate,L-carnitine, niacinamide, calcium pantothenate, cupric sulfate, vitaminA palmitate, thiamine chloride hydrochloride, riboflavin, pyridoxinehydrochloride, folic acid, manganese sulfate, potassium iodide,phylloquinone, biotin, sodium selenite, vitamin D3 and cyanocobalamin.

[0188] D. Isomil® 20 Soy Formula with Iron Ready to Feed, 20 Cal/fl oz.:

[0189] Usage: When a soy feeding is desired.

[0190] Ingredients: (Pareve) 85% water, 4.9% corn syrup, 2.6%sugar(sucrose), 2.1% soy oil, 1.9% soy protein isolate, 1.4% coconutoil, 0.15% calcium citrate, 0. 11% calcium phosphate tribasic, potassiumcitrate, potassium phosphate monobasic, potassium chloride, mono- anddiglycerides, soy lecithin, carrageenan, abscorbic acid, L-methionine,magnesium chloride, potassium phosphate dibasic, sodium chloride,choline chloride, taurine, ferrous sulfate, m-inositol, alpha-tocopherylacetate, zinc sulfate, L-carnitine, niacinamide, calcium pantothenate,cupric sulfate, vitamin A palmitate, thiamine chloride hydrochloride,riboflavin, pyridoxine hydrochloride, folic acid, manganese sulfate,potassium iodide, phylloquinone, biotin, sodium selenite, vitamin D3 andcyanocobalamin.

[0191] E. Similac® Infant Formula:

[0192] Usage: When an infant formula is needed: if the decision is madeto discontinue breastfeeding before age 1 year, if a supplement tobreastfeeding is needed or as a routine feeding if breastfeeding is notadopted.

[0193] Features:

[0194] Protein of appropriate quality and quantity for good growth;heat-denatured, which reduces the risk of milk-associated enteric bloodloss.

[0195] Fat from a blend of vegetable oils (doubly homogenized),providing essential linoleic acid that is easily absorbed.

[0196] Carbohydrate as lactose in proportion similar to that of humanmilk.

[0197] Low renal solute load to minimize stress on developing organs.

[0198] Powder, Concentrated Liquid and Ready To Feed forms. Ingredients:(-D) Water, nonfat milk, lactose, soy oil, coconut oil, mono- anddiglycerides, soy lecithin, abscorbic acid, carrageenan, cholinechloride, taurine, m-inositol, alpha-tocopheryl acetate, zinc sulfate,niacinamide, ferrous sulfate, calcium pantothenate, cupric sulfate,vitamin A palmitate, thiamine chloride hydrochloride, riboflavin,pyridoxine hydrochloride, folic acid, manganese sulfate, phylloquinone,biotin, sodium selenite, vitamin D3 and cyanocobalamin.

[0199] F. Similac® NeoCare Premature Infant Formula with Iron:

[0200] Usage: For premature infants' special nutritional needs afterhospital discharge. Similac NeoCare is a nutritionally complete formuladeveloped to provide premature infants with extra calories, protein,vitamins and minerals needed to promote catch-up growth and supportdevelopment.

[0201] Features:

[0202] Reduces the need for caloric and vitamin supplementation. Morecalories (22 Cal/fl oz) than standard term formulas (20 Cal/fl oz).

[0203] Highly absorbed fat blend, with medium-chain triglycerides (MCToil) to help meet the special digestive needs of premature infants.

[0204] Higher levels of protein, vitamins and minerals per 100 caloriesto extend the nutritional support initiated in-hospital.

[0205] More calcium and phosphorus for improved bone mineralization.

[0206] Ingredients: -D Corn syrup solids, nonfat milk, lactose, wheyprotein concentrate, soy oil, high-oleic safflower oil, fractionatedcoconut oil (medium chain triglycerides), coconut oil, potassiumcitrate, calcium phosphate tribasic, calcium carbonate, ascorbic acid,magnesium chloride, potassium chloride, sodium chloride, taurine,ferrous sulfate, m-inositol, choline chloride, ascorbyl palmitate,L-carnitine, alpha-tocopheryl acetate, zinc sulfate, niacinamide, mixedtocopherols, sodium citrate, calcium pantothenate, cupric sulfate,thiamine chloride hydrochloride, vitamin A palmitate, beta carotene,riboflavin, pyridoxine hydrochloride, folic acid, manganese sulfate,phylloquinone, biotin, sodium selenite, vitamin D3 and cyanocobalamin.

[0207] G. Similac Natural Care Low-Iron Human Milk Fortifier Ready touse, 24 Cal/fl oz.:

[0208] Usage: Designed to be mixed with human milk or to be fedalternatively with human milk to low-birth-weight infants.

[0209] Ingredients: -D Water, nonfat milk, hydrolyzed cornstarch,lactose, fractionated coconut oil (medium-chain triglycerides), wheyprotein concentrate, soy oil, coconut oil, calcium phosphate tribasic,potassium citrate, magnesium chloride, sodium citrate, ascorbic acid,calcium carbonate, mono and diglycerides, soy lecithin, carrageenan,choline chloride, m-inositol, taurine, niacinamide, L-carnitine, alphatocopheryl acetate, zinc sulfate, potassium chloride, calciumpantothenate, ferrous sulfate, cupric sulfate, riboflavin, vitamin Apalmitate, thiamine chloride hydrochloride, pyridoxine hydrochloride,biotin, folic acid, manganese sulfate, phylloquinone, vitamin D3, sodiumselenite and cyanocobalamin.

[0210] Various PUFAs of this invention can be substituted and/or addedto the infant formulae described above and to other infant formulaeknown to those in the art.

[0211] II. Nutritional Formulations

[0212] A. ENSURE®

[0213] Usage: ENSURE is a low-residue liquid food designed primarily asan oral nutritional supplement to be used with or between meals or, inappropriate amounts, as a meal replacement. ENSURE is lactose- andgluten-free, and is suitable for use in modified diets, includinglow-cholesterol diets. Although it is primarily an oral supplement, itcan be fed by tube.

[0214] Patient Conditions:

[0215] For patients on modified diets

[0216] For elderly patients at nutrition risk

[0217] For patients with involuntary weight loss

[0218] For patients recovering from illness or surgery

[0219] For patients who need a low-residue diet

[0220] Ingredients: -D Water, Sugar (Sucrose), Maltodextrin (Corn),Calcium and Sodium Caseinates, High-Oleic Safflower Oil, Soy ProteinIsolate, Soy Oil, Canola Oil, Potassium Citrate, Calcium PhosphateTribasic, Sodium Citrate, Magnesium Chloride, Magnesium PhosphateDibasic, Artificial Flavor, Sodium Chloride, Soy Lecithin, CholineChloride, Ascorbic Acid, Carrageenan, Zinc Sulfate, Ferrous Sulfate,Alpha-Tocopheryl Acetate, Gellan Gum, Niacinamide, Calcium Pantothenate,Manganese Sulfate, Cupric Sulfate, Vitamin A Palmitate, ThiamineChloride Hydrochloride, Pyridoxine Hydrochloride, Riboflavin, FolicAcid, Sodium Molybdate, Chromium Chloride, Biotin, Potassium Iodide,Sodium Selenate.

[0221] B. ENSURE® BARS:

[0222] Usage: ENSURE BARS are complete, balanced nutrition forsupplemental use between or with meals. They provide a delicious,nutrient-rich alternative to other snacks. ENSURE BARS contain <1 glactose/bar, and Chocolate Fudge Brownie flavor is gluten-free. (HoneyGraham Crunch flavor contains gluten.)

[0223] Patient Conditions:

[0224] For patients who need extra calories, protein, vitamins andminerals.

[0225] Especially useful for people who do not take in enough caloriesand nutrients.

[0226] For people who have the ability to chew and swallow

[0227] Not to be used by anyone with a peanut allergy or any type ofallergy to nuts.

[0228] Ingredients: Honey Graham Crunch—High-Fructose Corn Syrup, SoyProtein Isolate, Brown Sugar, Honey, Maltodextrin (Corn), Crisp Rice(Milled Rice, Sugar [Sucrose], Salt [Sodium Chloride] and Malt), OatBran, Partially Hydrogenated Cottonseed and Soy Oils, SoyPolysaccharide, Glycerine, Whey Protein Concentrate, Polydextrose,Fructose, Calcium Caseinate, Cocoa Powder, Artificial Flavors, CanolaOil, High-Oleic Safflower Oil, Nonfat Dry Milk, Whey Powder, SoyLecithin and Corn Oil. Manufactured in a facility that processes nuts.

[0229] Vitamins and Minerals: Calcium Phosphate Tribasic, PotassiumPhosphate Dibasic, Magnesium Oxide, Salt (Sodium Chloride), PotassiumChloride, Ascorbic Acid, Ferric Orthophosphate, Alpha-TocopherylAcetate, Niacinamide, Zinc Oxide, Calcium Pantothenate, CopperGluconate, Manganese Sulfate, Riboflavin, Beta Carotene, PyridoxineHydrochloride, Thiamine Mononitrate, Folic Acid, Biotin, ChromiumChloride, Potassium Iodide, Sodium Selenate, Sodium Molybdate,Phylloquinone, Vitamin D3 and Cyanocobalamin.

[0230] Protein: Honey Graham Crunch—The protein source is a blend of soyprotein isolate and milk proteins. Soy protein isolate 74% Milk proteins26%

[0231] Fat: Honey Graham Crunch—The fat source is a blend of partiallyhydrogenated cottonseed and soybean, canola, high oleic safflower, oils,and soy lecithin. Partially hydrogenated cottonseed and soybean oil 76% Canola oil 8% High-oleic safflower oil 8% Corn oil 4% Soy lecithin 4%

[0232] Carbohydrate: Honey Graham Crunch—The carbohydrate source is acombination of high-fructose corn syrup, brown sugar, maltodextrin,honey, crisp rice, glycerine, soy polysaccharide, and oat bran.High-fructose corn syrup 24% Brown sugar 21% Maltodextrin 12% Honey 11%Crisp rice  9% Glycerine  9% Soy Polysaccharide  7% Oat bran  7%

[0233] C. ENSURE® HIGH PROTEIN:

[0234] Usage: ENSURE HIGH PROTEIN is a concentrated, high-protein liquidfood designed for people who require additional calories, protein,vitamins, and minerals in their diets. It can be used as an oralnutritional supplement with or between meals or, in appropriate amounts,as a meal replacement. ENSURE HIGH PROTEIN is lactose- and gluten-free,and is suitable for use by people recovering from general surgery or hipfractures and by patients at risk for pressure ulcers.

[0235] Patient Conditions:

[0236] For patients who require additional calories, protein, vitamins,and minerals, such as patients recovering from general surgery or hipfractures, patients at risk for pressure ulcers, and patients onlow-cholesterol diets

[0237] Features:

[0238] Low in saturated fat

[0239] Contains 6 g of total fat and <5 mg of cholesterol per serving

[0240] Rich, creamy taste

[0241] Excellent source of protein, calcium, and other essentialvitamins and minerals

[0242] For low-cholesterol diets

[0243] Lactose-free, easily digested

[0244] Ingredients:

[0245] Vanilla Supreme: -D Water, Sugar (Sucrose), Maltodextrin (Corn),Calcium and Sodium Caseinates, High-OIeic Safflower Oil, Soy ProteinIsolate, Soy Oil, Canola Oil, Potassium Citrate, Calcium PhosphateTribasic, Sodium Citrate, Magnesium Chloride, Magnesium PhosphateDibasic, Artificial Flavor, Sodium Chloride, Soy Lecithin, CholineChloride, Ascorbic Acid, Carrageenan, Zinc Sulfate, Ferrous Suffate,Alpha-Tocopheryl Acetate, Gellan Gum, Niacinamide, Calcium Pantothenate,Manganese Sulfate, Cupric Sulfate, Vitamin A Palmitate, ThiamineChloride Hydrochloride, Pyridoxine Hydrochloride, Riboflavin, FolicAcid, Sodium Molybdate, Chromium Chloride, Biotin, Potassium Iodide,Sodium Selenate, Phylloquinone, Vitamin D3 and Cyanocobalamin.

[0246] Protein:

[0247] The protein source is a blend of two high-biologic-valueproteins: casein and soy. Sodium and calcium caseinates 85% Soy proteinisolate 15%

[0248] Fat:

[0249] The fat source is a blend of three oils: high-oleic safflower,canola, and soy. High-oleic safflower oil 40% Canola oil 30% Soy oil 30%

[0250] The level of fat in ENSURE HIGH PROTEIN meets American HeartAssociation (AHA) guidelines. The 6 grams of fat in ENSURE HIGH PROTEINrepresent 24% of the total calories, with 2.6% of the fat being fromsaturated fatty acids and 7.9% from polyunsaturated fatty acids. Thesevalues are within the AHA guidelines of <30% of total calories from fat,<10% of the calories from saturated fatty acids, and <10% of totalcalories from polyunsaturated fatty acids.

[0251] Carbohydrate:

[0252] ENSURE HIGH PROTEIN contains a combination of maltodextrin andsucrose. The mild sweetness and flavor variety (vanilla supreme,chocolate royal, wild berry, and banana), plus VARI-FLAVORS® Flavor Pacsin pecan, cherry, strawberry, lemon, and orange, help to prevent flavorfatigue and aid in patient compliance.

[0253] Vanilla and Other Nonchocolate Flavors: Sucrose 60% Maltodextrin40% Chocolate: Sucrose 70% Maltodextrin 30%

[0254] D. ENSURE® LIGHT

[0255] Usage: ENSURE LIGHT is a low-fat liquid food designed for use asan oral nutritional supplement with or between meals. ENSURE LIGHT islactose- and gluten-free, and is suitable for use in modified diets,including low-cholesterol diets.

[0256] Patient Conditions:

[0257] For normal-weight or overweight patients who need extra nutritionin a supplement that contains 50% less fat and 20% fewer calories thanENSURE.

[0258] For healthy adults who don't eat right and need extra nutrition.

[0259] Features:

[0260] Low in fat and saturated fat

[0261] Contains 3 g of total fat per serving and <5 mg cholesterol

[0262] Rich, creamy taste

[0263] Excellent source of calcium and other essential vitamins andminerals

[0264] For low-cholesterol diets

[0265] Lactose-free, easily digested

[0266] Ingredients:

[0267] French Vanilla: -D Water, Maltodextrin (Corn), Sugar (Sucrose),Calcium Caseinate, High-Oleic Safflower Oil, Canola Oil, MagnesiumChloride, Sodium Citrate, Potassium Citrate, Potassium PhosphateDibasic, Magnesium Phosphate Dibasic, Natural and Artificial Flavor,Calcium Phosphate Tribasic, Cellulose Gel, Choline Chloride, SoyLecithin, Carrageenan, Salt (Sodium Chloride), Ascorbic Acid, CelluloseGum, Ferrous Sulfate, Alpha-Tocopheryl Acetate, Zinc Sulfate,Niacinamide, Manganese Sulfate, Calcium Pantothenate, Cupric Sulfate,Thiamine Chloride Hydrochloride, Vitamin A Palmitate, PyridoxineHydrochloride, Riboflavin, Chromium Chloride, Folic Acid, SodiumMolybdate, Biotin, Potassium Iodide, Sodium Selenate, Phylloquinone,Vitamin D3 and Cyanocobalamin.

[0268] Protein:

[0269] The protein source is calcium caseinate. Calcium caseinate 100%

[0270] Fat:

[0271] The fat source is a blend of two oils: high-oleic safflower andcanola. High-oleic safflower oil 70% Canola oil 30%

[0272] The level of fat in ENSURE LIGHT meets American Heart Association(AHA) guidelines. The 3 grams of fat in ENSURE LIGHT represent 13.5% ofthe total calories, with 1.4% of the fat being from saturated fattyacids and 2.6% from polyunsaturated fatty acids. These values are withinthe AHA guidelines of <30% of total calories from fat, <10% of the,calories from saturated fatty acids, and <10% of total calories frompolyunsaturated fatty acids.

[0273] Carbohydrate:

[0274] ENSURE LIGHT contains a combination of maltodextrin and sucrose.The chocolate flavor contains corn syrup as well. The mild sweetness andflavor variety (French vanilla, chocolate supreme, strawberry swirl),plus VARI-FLAVORS® Flavor Pacs in pecan, cherry, strawberry, lemon, andorange, help to prevent flavor fatigue and aid in patient compliance.

[0275] Vanilla and Other Nonchocolate Flavors: Sucrose   51%Maltodextrin   49% Chocolate: Sucrose 47.0% Corn Syrup 26.5%Maltodextrin 26.5%

[0276] Vitamins and Minerals:

[0277] An 8-fl-oz serving of ENSURE LIGHT provides at least 25% of theRDIs for 24 key vitamins and minerals.

[0278] Caffeine:

[0279] Chocolate flavor contains 2.1 mg caffeine/8 fl oz.

[0280] E. ENSURE PLUS®

[0281] Usage: ENSURE PLUS is a high-calorie, low-residue liquid food foruse when extra calories and nutrients, but a normal concentration ofprotein, are needed. It is designed primarily as an oral nutritionalsupplement to be used with or between meals or, in appropriate amounts,as a meal replacement. ENSURE PLUS is lactose- and gluten-free. Althoughit is primarily an oral nutritional supplement, it can be fed by tube.

[0282] Patient Conditions:

[0283] For patients who require extra calories and nutrients, but anormal concentration of protein, in a limited volume

[0284] For patients who need to gain or maintain healthy weight

[0285] Features:

[0286] Rich, creamy taste

[0287] Good source of essential vitamins and minerals

[0288] Ingredients:

[0289] Vanilla: -D Water, Corn Syrup, Maltodextrin (Corn), Corn Oil,Sodium and Calcium Caseinates, Sugar (Sucrose), Soy Protein Isolate,Magnesium Chloride, Potassium Citrate, Calcium Phosphate Tribasic, SoyLecithin, Natural and Artificial Flavor, Sodium Citrate, PotassiumChloride, Choline Chloride, Ascorbic Acid, Carrageenan, Zinc Sulfate,Ferrous Sulfate, Alpha-Tocopheryl Acetate, Niacinamide, CalciumPantothenate, Manganese Sulfate, Cupric Sulfate, Thiamine ChlorideHydrochloride, Pyridoxine Hydrochloride, Riboflavin, Vitamin APalmitate, Folic Acid, Biotin, Chromium Chloride, Sodium Molybdate,Potassium Iodide, Sodium Selenite, Phylloquinone, Cyanocobalamin andVitamin D3.

[0290] Protein:

[0291] The protein source is a blend of two high-biologic-valueproteins: casein and soy. Sodium and calcium caseinates 84% Soy proteinisolate 16%

[0292] Fat:

[0293] The fat source is corn oil. Corn oil 100%

[0294] Carbohydrate:

[0295] ENSURE PLUS contains a combination of maltodextrin and sucrose.The mild sweetness and flavor variety (vanilla, chocolate, strawberry,coffee, buffer pecan, and eggnog), plus VARI-FLAVORS® Flavor Pacs inpecan, cherry, strawberry, lemon, and orange, help to prevent flavorfatigue and aid in patient compliance.

[0296] Vanilla, Strawberry, Butter Pecan, and Coffee Flavors: Corn Syrup39% Maltodextrin 38% Sucrose 23% Chocolate and eggnog flavors: CornSyrup 36% Maltodextrin 34% Sucrose 30%

[0297] Vitamins and Minerals:

[0298] An 8-fl-oz serving of ENSURE PLUS provides at least 15% of theRDIs for 25 key Vitamins and minerals.

[0299] Caffeine:

[0300] Chocolate flavor contains 3.1 mg Caffeine/8 fl oz. Coffee flavorcontains a trace amount of caffeine.

[0301] F. ENSURE PLUS® HN

[0302] Usage: ENSURE PLUS HN is a nutritionally complete high-calorie,high-nitrogen liquid food designed for people with higher calorie andprotein needs or limited volume tolerance. It may be used for oralsupplementation or for total nutritional support by tube. ENSURE PLUS HNis lactose- and gluten-free.

[0303] Patient Conditions:

[0304] For patients with increased calorie and protein needs, such asfollowing surgery or injury.

[0305] For patients with limited volume tolerance and early satiety.

[0306] Features:

[0307] For supplemental or total nutrition

[0308] For oral or tube feeding

[0309] 1.5 CaVmL,

[0310] High nitrogen

[0311] Calorically dense

[0312] Ingredients:

[0313] Vanilla: -D Water, Maltodextrin (Corn), Sodium and CalciumCaseinates, Corn Oil, Sugar (Sucrose), Soy Protein Isolate, MagnesiumChloride, Potassium Citrate, Calcium Phosphate Tribasic, Soy Lecithin,Natural and Artificial Flavor, Sodium Citrate, Choline Chloride,Ascorbic Acid, Taurine, L-Carnitine, Zinc Sulfate, Ferrous Sulfate,Alpha-Tocopheryl Acetate, Niacinamide, Carrageenan, CalciumPantothenate, Manganese Sulfate, Cupric Sulfate, Thiamine ChlorideHydrochloride, Pyridoxine Hydrochloride, Riboflavin, Vitamin APalmitate, Folic Acid, Biotin, Chromium Chloride, Sodium Molybdate,Potassium Iodide, Sodium Selenite, Phylloquinone, Cyanocobalamin andVitamin D3.

[0314] G. ENSURE® POWDER:

[0315] Usage: ENSURE POWDER (reconstituted with water) is a low-residueliquid food designed primarily as an oral nutritional supplement to beused with or between meals. ENSURE POWDER is lactose- and gluten-free,and is suitable for use in modified diets, including low-cholesteroldiets.

[0316] Patient Conditions:

[0317] For patients on modified diets

[0318] For elderly patients at nutrition risk

[0319] For patients recovering from illness/surgery

[0320] For patients who need a low-residue diet

[0321] Features:

[0322] Convenient, easy to mix

[0323] Low in saturated fat

[0324] Contains 9 g of total fat and <5 mg of cholesterol per serving

[0325] High in vitamins and minerals

[0326] For low-cholesterol diets

[0327] Lactose-free, easily digested

[0328] Ingredients: -D Corn Syrup, Maltodextrin (Corn), Sugar (Sucrose),Corn Oil, Sodium and Calcium Caseinates, Soy Protein Isolate, ArtificialFlavor, Potassium Citrate, Magnesium Chloride, Sodium Citrate, CalciumPhosphate Tribasic, Potassium Chloride, Soy Lecithin, Ascorbic Acid,Choline Chloride, Zinc Sulfate, Ferrous Sulfate, Alpha-TocopherylAcetate, Niacinamide, Calcium Pantothenate, Manganese Sulfate, ThiamineChloride Hydrochloride, Cupric Sulfate, Pyridoxine Hydrochloride,Riboflavin, Vitamin A Palmitate, Folic Acid, Biotin, Sodium Molybdate,Chromium Chloride, Potassium Iodide, Sodium Selenate, Phylloquinone,Vitamin D3 and Cyanocobalamin.

[0329] Protein:

[0330] The protein source is a blend of two high-biologic-valueproteins: casein and soy. Sodium and calcium caseinates 84% Soy proteinisolate 16%

[0331] Fat:

[0332] The fat source is corn oil. Corn oil 100%

[0333] Carbohydrate:

[0334] ENSURE POWDER contains a combination of corn syrup, maltodextrin,and sucrose. The mild sweetness of ENSURE POWDER, plus VARI-FLAVORS®Flavor Pacs in pecan, cherry, strawberry, lemon, and orange, helps toprevent flavor fatigue and aid in patient compliance. Vanilla: CornSyrup 35% Maltodextrin 35% Sucrose 30%

[0335] H. ENSURE® PUDDING

[0336] Usage: ENSURE PUDDING is a nutrient-dense supplement providingbalanced nutrition in a nonliquid form to be used with or between meals.It is appropriate for consistency-modified diets (e.g., soft, pureed, orfull liquid) or for people with swallowing impairments. ENSURE PUDDINGis gluten-free.

[0337] Patient Conditions:

[0338] For patients on consistency-modified diets (e.g., soft, pureed,or full liquid)

[0339] For patients with swallowing impairments

[0340] Features:

[0341] Rich and creamy, good taste

[0342] Good source of essential vitamins and minerals

[0343] Convenient-needs no refrigeration

[0344] Gluten-free

[0345] Nutrient Profile per 5 oz: Calories 250, Protein 10.9%, Total Fat34.9%, Carbohydrate 54.2%

[0346] Ingredients:

[0347] Vanilla: -D Nonfat Milk, Water, Sugar (Sucrose), PartiallyHydrogenated Soybean Oil, Modified Food Starch, Magnesium Sulfate,Sodium Stearoyl Lactylate, Sodium Phosphate Dibasic, Artificial Flavor,Ascorbic Acid, Zinc Sulfate, Ferrous Sulfate, Alpha-Tocopheryl Acetate,Choline Chloride, Niacinamide, Manganese Sulfate, Calcium Pantothenate,FD&C Yellow #5, Potassium Citrate, Cupric Sulfate, Vitamin A Palmitate,Thiamine Chloride Hydrochloride, Pyridoxine Hydrochloride, Riboflavin,FD&C Yellow #6, Folic Acid, Biotin, Phylloquinone, Vitamin D3 andCyanocobalamin.

[0348] Protein:

[0349] The protein source is nonfat milk. Nonfat milk 100%

[0350] Fat:

[0351] The fat source is hydrogenated soybean oil. Hydrogenated soybeanoil 100%

[0352] Carbohydrate:

[0353] ENSURE PUDDING contains a combination of sucrose and modifiedfood starch. The mild sweetness and flavor variety (vanilla, chocolate,butterscotch, and tapioca) help prevent flavor fatigue. The productcontains 9.2 grams of lactose per serving.

[0354] Vanilla and Other Nonchocolate Flavors: Sucrose 56% Lactose 27%Modified food starch 17% Chocolate: Sucrose 58% Lactose 26% Modifiedfood starch 16%

[0355] I. ENSURE® WITH FIBER:

[0356] Usage: ENSURE WITH FIBER is a fiber-containing, nutritionallycomplete liquid food designed for people who can benefit from increaseddietary fiber and nutrients. ENSURE WITH FIBER is suitable for peoplewho do not require a low-residue diet. It can be fed orally or by tube,and can be used as a nutritional supplement to a regular diet or, inappropriate amounts, as a meal replacement. ENSURE WITH FIBER islactose- and gluten-free, and is suitable for use in modified diets,including low-cholesterol diets.

[0357] Patient Conditions:

[0358] For patients who can benefit from increased dietary fiber andnutrients

[0359] Features:

[0360] New advanced formula-low in saturated fat, higher in vitamins andminerals

[0361] Contains 6 g of total fat and <5 mg of cholesterol per serving

[0362] Rich, creamy taste

[0363] Good source of fiber

[0364] Excellent source of essential vitamins and minerals

[0365] For low-cholesterol diets

[0366] Lactose- and gluten-free

[0367] Ingredients:

[0368] Vanilla: -D Water; Maltodextrin (Corn), Sugar (Sucrose), Sodiumand Calcium Caseinates, Oat Fiber, High-Oleic Safflower Oil, Canola Oil,Soy Protein Isolate, Corn Oil, Soy Fiber, Calcium Phosphate Tribasic,Magnesium Chloride, Potassium Citrate, Cellulose Gel, Soy Lecithin,Potassium Phosphate Dibasic, Sodium Citrate, Natural and ArtificialFlavors, Choline Chloride, Magnesium Phosphate, Ascorbic Acid, CelluloseGum, Potassium Chloride, Carrageenan, Ferrous Sulfate, Alpha-TocopherylAcetate, Zinc Sulfate, Niacinamide, Manganese Sulfate, CalciumPantothenate, Cupric Sulfate, Vitamin A Palmitate, Thiamine ChlorideHydrochloride, Pyridoxine Hydrochloride, Riboflavin, Folic Acid,Chromium Chloride, Biotin, Sodium Molybdate, Potassium Iodide, SodiumSelenate, Phylloquinone, Vitamin D3 and Cyanocobalamin.

[0369] Protein:

[0370] The protein source is a blend of two high-biologic-valueproteins-casein and soy.

[0371] Sodium and calcium caseinates

[0372] 80%

[0373] Soy protein isolate

[0374] 20%

[0375] Fat:

[0376] The fat source is a blend of three oils: high-oleic safflower,canola, and corn.

[0377] High-oleic safflower oil

[0378] 40%

[0379] Canola oil

[0380] 40%

[0381] Corn oil

[0382] 20%

[0383] The level of fat in ENSURE WITH FIBER meets American HeartAssociation (AHA) guidelines. The 6 grams of fat in ENSURE WITH FIBERrepresent 22% of the total calories, with 2.01% of the fat being fromsaturated fatty acids and 6.7% from polyunsaturated fatty acids. Thesevalues are within the AHA guidelines of ≦30% of total calories from fat,<10% of the calories from saturated fatty acids, and ≦10% of totalcalories from polyunsaturated fatty acids.

[0384] Carbohydrate:

[0385] ENSURE WITH FIBER contains a combination of maltodextrin andsucrose. The mild sweetness and flavor variety (vanilla, chocolate, andbutter pecan), plus VARI-FLAVORS® Flavor Pacs in pecan, cherry,strawberry, lemon, and orange, help to prevent flavor fatigue and aid inpatient compliance. Vanilla and other nonchocolate flavors: Maltodextrin66% Sucrose 25% Oat Fiber  7% Soy Fiber  2% Chocolate: Maltodextrin 55%Sucrose 36% Oat Fiber  7% Soy Fiber  2%

[0386] Fiber:

[0387] The fiber blend used in ENSURE WITH FIBER consists of oat fiberand soy polysaccharide. This blend results in approximately 4 grams oftotal dietary fiber per 8-fl. oz can. The ratio of insoluble to solublefiber is 95:5.

[0388] The various nutritional supplements described above and known toothers of skill in the art can be substituted and/or supplemented withthe PUFAs produced in accordance with the present invention.

[0389] J. Oxepa™ Nutritional Product

[0390] Oxepa is a low-carbohydrate, calorically dense, enteralnutritional product designed for the dietary management of patients withor at risk for ARDS. It has a unique combination of ingredients,including a patented oil blend containing eicosapentaenoic acid (EPAfrom fish oil), γ-linolenic acid (GLA from borage oil), and elevatedantioxidant levels.

[0391] Caloric Distribution:

[0392] Caloric density is high at 1.5 Cal/mL (355 Cal/8 fl oz), tominimize the volume required to meet energy needs.

[0393] The distribution of Calories in Oxepa is shown in Table IV. TABLEIV Caloric Distribution of Oxepa per 8 fl oz. per liter % of CalCalories 355 1,500 — Fat (g) 22.2 93.7 55.2 Carbohydrate (g) 25 105.528.1 Protein (g) 14.8 62.5 16.7 Water (g) 186 785 —

[0394] Fat:

[0395] Oxepa contains 22.2 g of fat per 8-fl oz serving (93.7 g/L).

[0396] The fat source is an oil blend of 31.8% canola oil, 25%medium-chain triglycerides (MCTs), 20% borage oil, 20% fish oil, and3.2% soy lecithin. The typical fatty acid profile of Oxepa is shown inTable V.

[0397] Oxepa provides a balanced amount of polyunsaturated,monounsaturated, and saturated fatty acids, as shown in Table VI.

[0398] -Medium-chain trigylcerides (MCTs)—25% of the fat blend—aidgastric emptying because they are absorbed by the intestinal tractwithout emulsification by bile acids.

[0399] The various fatty acid components of Oxepa™ nutritional productcan be substituted and/or supplemented with the PUFAs produced inaccordance with this invention. TABLE V Typical Fatty Acid Profile %Total Fatty Acids g/8 fl oz* 9/L* Caproic (6:0) 0.2 0.04 0.18 Caprylic(8:0) 14.69 3.1 13.07 Capric (10:0) 11.06 2.33 9.87 Palmitic (16:0) 5.591.18 4.98 Palmitoleic 1.82 0.38 1.62 Stearic 1.94 0.39 1.64 Oleic 24.445.16 21.75 Linoleic 16.28 3.44 14.49 α-Linolenic 3.47 0.73 3.09γ-Linolenic 4.82 1.02 4.29 Eicosapentaenoic 5.11 1.08 4.55n-3-Docosapent- 0.55 0.12 0.49 aenoic Docosahexaenoic 2.27 0.48 2.02Others 7.55 1.52 6.72

[0400] Fatty acids equal approximately 95% of total fat. TABLE VI FatProfile of Oxepa. % of total calories from fat 55.2 Polyunsaturatedfatty acids 31.44 g/L Monounsaturated fatty acids 25.53 g/L Saturatedfatty acids 32.38 g/L n-6 to n-3 ratio 1.75:1 Cholesterol 9.49 mg/8 floz 40.1 mg/L

[0401] Carbohydrate:

[0402] The carbohydrate content is 25.0 g per 8-fl-oz serving (105.5g/L).

[0403] The carbohydrate sources are 45% maltodextrin (a complexcarbohydrate) and 55% sucrose (a simple sugar), both of which arereadily digested and absorbed.

[0404] The high-fat and low-carbohydrate content of Oxepa is designed tominimize carbon dioxide (CO2) production. High CO2 levels can complicateweaning in ventilator-dependent patients. The low level of carbohydratealso may be useful for those patients who have developed stress-inducedhyperglycemia.

[0405] Oxepa is lactose-free.

[0406] Dietary carbohydrate, the amino acids from protein, and theglycerol moiety of fats can be converted to glucose within the body.Throughout this process, the carbohydrate requirements ofglucose-dependent tissues (such as the central nervous system and redblood cells) are met. However, a diet free of carbohydrates can lead toketosis, excessive catabolism of tissue protein, and loss of fluid andelectrolytes. These effects can be prevented by daily ingestion of 50 to100 g of digestible carbohydrate, if caloric intake is adequate. Thecarbohydrate level in Oxepa is also sufficient to minimizegluconeogenesis, if energy needs are being met.

[0407] Protein:

[0408] Oxepa contains 14.8 g of protein per 8-fl-oz serving (62.5 g/L).

[0409] The total calorie/nitrogen ratio (150:1) meets the need ofstressed patients.

[0410] Oxepa provides enough protein to promote anabolism and themaintenance of lean body mass without precipitating respiratoryproblems. High protein intakes are a concern in patients withrespiratory insufficiency. Although protein has little effect on CO2production, a high protein diet will increase ventilatory drive.

[0411] The protein sources of Oxepa are 86.8% sodium caseinate and 13.2%calcium caseinate.

[0412] The amino acid profile of the protein system in Oxepa meets orsurpasses the standard for high quality protein set by the NationalAcademy of Sciences.

[0413] Oxepa is gluten-free.

1 54 1 1335 DNA Homo sapiens 1 atggcccccg acccggtggc cgccgagaccgcggctcagg gacctacccc gcgctacttc 60 acctgggacg aggtggccca gcgctcagggtgcgaggagc ggtggctagt gatcgaccgt 120 aaggtgtaca acatcagcga gttcacccgccggcatccag ggggctcccg ggtcatcagc 180 cactacgccg ggcaggatgc cacggatccctttgtggcct tccacatcaa caagggcctt 240 gtgaagaagt atatgaactc tctcctgattggagaactgt ctccagagca gcccagcttt 300 gagcccacca agaataaaga gctgacagatgagttccggg agctgcgggc cacagtggag 360 cggatggggc tcatgaaggc caaccatgtcttcttcctgc tgtacctgct gcacatcttg 420 ctgctggatg gtgcagcctg gctcaccctttgggtctttg ggacgtcctt tttgcccttc 480 ctcctctgtg cggtgctgct cagtgcagttcaggcccagg ctggctggct gcagcatgac 540 tttgggcacc tgtcggtctt cagcacctcaaagtggaacc atctgctaca tcattttgtg 600 attggccacc tgaagggggc ccccgccagttggtggaacc acatgcactt ccagcaccat 660 gccaagccca actgcttccg caaagacccagacatcaaca tgcatccctt cttctttgcc 720 ttggggaaga tcctctctgt ggagcttgggaaacagaaga aaaaatatat gccgtacaac 780 caccagcaca aatacttctt cctaattgggcccccagcct tgctgcctct ctacttccag 840 tggtatattt tctattttgt tatccagcgaaagaagtggg tggacttggc ctggatgatt 900 accttctacg tccgcttctt cctcacttatgtgccactat tggggctgaa agccttcctg 960 ggccttttct tcatagtcag gttcctggaaagcaactggt ttgtgtgggt gacacagatg 1020 aaccatattc ccatgcacat tgatcatgaccggaacatgg actgggtttc cacccagctc 1080 ctggccacat gcaatgtcca caagtctgccttcaatgact ggttcagtgg acacctcaac 1140 ttccagattg agcaccatct ttttcccacgatgcctcgac acaattacca caaagtggct 1200 cccctggtgc agtccttgtg tgccaagcgtggcatagagt accagtccaa gcccctgctg 1260 tcagccttcg ccgacatcat ccactcactaaaggagtcag ggcagctctg gctagatgcc 1320 tatcttcacc aataa 1335 2 1219 DNAHomo sapiens 2 gcacgccgac cggcgccggg agatcctggc aaagtatcca gagataaagtccttgatgaa 60 acctgatccc aatttgatat ggattataat tatgatggtt ctcacccagttgggtgcatt 120 ttacatagta aaagacttgg actggaaatg ggtcatattt ggggcctatgcgtttggcag 180 ttgcattaac cactcaatga ctctggctat tcatgagatt gcccacaatgctgcctttgg 240 caactgcaaa gcaatgtgga atcgctggtt tggaatgttt gctaatcttcctattgggat 300 tccatattca atttccttta agaggtatca catggatcat catcggtaccttggagctga 360 tggcgtcgat gtagatattc ctaccgattt tgagggctgg ttcttctgtaccgctttcag 420 aaagtttata tgggttattc ttcagcctct cttttatgcc tttcgacctctgttcatcaa 480 ccccaaacca attacgtatc tggaagttat caataccgtg gcacaggtcacttttgacat 540 tttaatttat tactttttgg gaattaaatc cttagtctac atgttggcagcatctttact 600 tggcctgggt ttgcacccaa tttctggaca ttttatagct gagcattacatgttcttaaa 660 gggtcatgaa acttactcat attatgggcc tctgaattta cttaccttcaatgtgggtta 720 tcataatgaa catcatgatt tccccaacat tcctggaaaa agtcttccactggtgaggaa 780 aatagcagct gaatactatg acaacctccc tcactacaat tcctggataaaagtactgta 840 tgattttgtg atggatgata caataagtcc ctactcaaga atgaagaggcaccaaaaagg 900 agagatggtg ctggagtaaa tatcattagt gccaaaggga ttcttctccaaaactttaga 960 tgataaaatg gaatttttgc attattaaac ttgagaccag tgatgctcagaagctcccct 1020 ggcacaattt cagagtaaga gctcggtgat accaagaagt gaatctggcttttaaacagt 1080 cagcctgact ctgtactgct cagtttcact cacaggaaac ttgtgacttgtgtattatcg 1140 tcattgagga tgtttcactc atgtctgtca ttttataagc atatcatttaaaaagcttct 1200 aaaaagctat ttcgccagg 1219 3 655 DNA Homo sapiens 3ttaccttcta cgtccgcttc ttcctcactt atgtgccact attggggctg aaagcttcct 60gggccttttc ttcatagtca ggttcctgga aagcaactgg tttgtgtggg tgacacagat 120gaaccatatt cccatgcaca ttgatcatga ccggaacatg gactgggttt ccacccagct 180ccaggccaca tgcaatgtcc acaagtctgc cttcaatgac tggttcagtg gacacctcaa 240cttccagatt gagcaccatc tttttcccac gatgcctcga cacaattacc acaaagtggc 300tcccctggtg cagtccttgt gtgccaagca tggcatagag taccagtcca agcccctgct 360gtcagccttc gccgacatca tccactcact aaaggagtca gggcagctct ggctagatgc 420ctatcttcac caataacaac agccaccctg cccagtctgg aagaagagga ggaagactct 480ggagccaagg cagaggggag cttgagggac aatgccacta tagtttaata ctcagagggg 540gttgggtttg gggacataaa gcctctgact caaactcctc ccttttatct tctagccaca 600gttctaagac ccaaagtggg gggtggacac agaagtccct aggagggaag gagct 655 4 304DNA Homo sapiens 4 gtcttttact ttggcaatgg ctggattcct accctcatcacggcctttgt ccttgctacc 60 tctcaggccc aagctggatg gctgcaacat gattatggccacctgtctgt ctacagaaaa 120 cccaagtgga accaccttgt ccacaaattc gtcattggccacttaaaggg tgcctctgcc 180 aactggtgga atcatcgcca cttccagcac cacgccaagcctaacatctt ccacaaggat 240 cccgatgtga acatgctgca cgtgtttgtt ctgggcgaatggcagcccat cgagtacggc 300 aaga 304 5 918 DNA Homo sapiens misc_feature(755)...(755) r = g or a at position 755 5 cagggaccta ccccgcgctacttcacctgg gacgaggtgg cccagcgctc agggtgcgag 60 gagcggtggc tagtgatcgaccgtaaggtg tacaacatca gcgagttcac ccgccggcat 120 ccagggggct cccgggtcatcagccactac gccgggcagg atgccacgga tccctttgtg 180 gccttccaca tcaacaagggccttgtgaag aagtatatga actctctcct gattggagaa 240 ctgtctccag agcagcccagctttgagccc accaagaata aagagctgac agatgagttc 300 cgggagctgc gggccacagtggagcggatg gggctcatga aggccaacca tgtcttcttc 360 ctgctgtacc tgctgcacatcttgctgctg gatggtgcag cctggctcac cctttgggtc 420 tttgggacgt cctttttgcccttcctcctc tgtgcggtgc tgctcagtgc agttcaggcc 480 caggctggct ggctgcagcatgactttggg cacctgtcgg tcttcagcac ctcaaagtgg 540 aaccatctgc tacatcattttgtgattggc cacctgaagg gggcccccgc cagttggtgg 600 aaccacatgc acttccagcaccatgccaag cccaactgct tccgcaaaga cccagacatc 660 aacatgcatc ccttcttctttgccttgggg aagatcctct ctgtggagct tgggaaacag 720 aagaaaaaat atatgccgtacaaccaccag cacaratact tcttcctaat tgggccccca 780 gccttgctgc ctctctacttccagtggtat attttctatt ttgttatcca gcgaaagaag 840 tgggtggact tggcctggatcagcaaacag gaatacgatg aagccgggct tccattgtcc 900 accgcaaatg cttctaaa 9186 1686 DNA Homo sapiens 6 gccacttaaa gggtgcctct gccaactggt ggaatcatcgccacttccag caccacgcca 60 agcctaacat cttccacaag gatcccgatg tgaacatgctgcacgtgttt gttctgggcg 120 aatggcagcc catcgagtac ggcaagaaga agctgaaatacctgccctac aatcaccagc 180 acgaatactt cttcctgatt gggccgccgc tgctcatccccatgtatttc cagtaccaga 240 tcatcatgac catgatcgtc cataagaact gggtggacctggcctgggcc gtcagctact 300 acatccggtt cttcatcacc tacatccctt tctacggcatcctgggagcc ctccttttcc 360 tcaacttcat caggttcctg gagagccact ggtttgtgtgggtcacacag atgaatcaca 420 tcgtcatgga gattgaccag gaggcctacc gtgactggttcagtagccag ctgacagcca 480 cctgcaacgt ggagcagtcc ttcttcaacg actggttcagtggacacctt aacttccaga 540 ttgagcacca cctcttcccc accatgcccc ggcacaacttacacaagatc gccccgctgg 600 tgaagtctct atgtgccaag catggcattg aataccaggagaagccgcta ctgagggccc 660 tgctggacat catcaggtcc ctgaagaagt ctgggaagctgtggctggac gcctaccttc 720 acaaatgaag ccacagcccc cgggacaccg tggggaaggggtgcaggtgg ggtgatggcc 780 agaggaatga tgggcttttg ttctgagggg tgtccgagaggctggtgtat gcactgctca 840 cggaccccat gttggatctt tctccctttc tcctctcctttttctcttca catctccccc 900 atagcaccct gccctcatgg gacctgccct ccctcagccgtcagccatca gccatggccc 960 tcccagtgcc tcctagcccc ttcttccaag gagcagagaggtggccaccg ggggtggctc 1020 tgtcctacct ccactctctg cccctaaaga tgggaggagaccagcggtcc atgggtctgg 1080 cctgtgagtc tccccttgca gcctggtcac taggcatcacccccgctttg gttcttcaga 1140 tgctcttggg gttcataggg gcaggtccta gtcgggcagggcccctgacc ctcccggcct 1200 ggcttcactc tccctgacgg ctgccattgg tccaccctttcatagagagg cctgctttgt 1260 tacaaagctc gggtctccct cctgcagctc ggttaagtacccgaggcctc tcttaagatg 1320 tccagggccc caggcccgcg ggcacagcca gcccaaaccttgggccctgg aagagtcctc 1380 caccccatca ctagagtgct ctgaccctgg gctttcacgggccccattcc accgcctccc 1440 caacttgagc ctgtgacctt gggaccaaag ggggagtccctcgtctcttg tgactcagca 1500 gaggcagtgg ccacgttcag ggaggggccg gctggcctggaggctcagcc caccctccag 1560 cttttcctca gggtgtcctg aggtccaaga ttctggagcaatctgaccct tctccaaagg 1620 ctctgttatc agctgggcag tgccagccaa tccctggccatttggcccca ggggacgtgg 1680 gccctg 1686 7 1843 DNA Homo sapiens 7gtcttttact ttggcaatgg ctggattcct accctcatca cggcctttgt ccttgctacc 60tctcaggccc aagctggatg gctgcaacat gattatggcc acctgtctgt ctacagaaaa 120cccaagtgga accaccttgt ccacaaattc gtcattggcc acttaaaggg tgcctctgcc 180aactggtgga atcatcgcca cttccagcac cacgccaagc ctaacatctt ccacaaggat 240cccgatgtga acatgctgca cgtgtttgtt ctgggcgaat ggcagcccat cgagtacggc 300aagaagaagc tgaaatacct gccctacaat caccagcacg aatacttctt cctgattggg 360ccgccgctgc tcatccccat gtatttccag taccagatca tcatgaccat gatcgtccat 420aagaactggg tggacctggc ctgggccgtc agctactaca tccggttctt catcacctac 480atccctttct acggcatcct gggagccctc cttttcctca acttcatcag gttcctggag 540agccactggt ttgtgtgggt cacacagatg aatcacatcg tcatggagat tgaccaggag 600gcctaccgtg actggttcag tagccagctg acagccacct gcaacgtgga gcagtccttc 660ttcaacgact ggttcagtgg acaccttaac ttccagattg agcaccacct cttccccacc 720atgccccggc acaacttaca caagatcgcc ccgctggtga agtctctatg tgccaagcat 780ggcattgaat accaggagaa gccgctactg agggccctgc tggacatcat caggtccctg 840aagaagtctg ggaagctgtg gctggacgcc taccttcaca aatgaagcca cagcccccgg 900gacaccgtgg ggaaggggtg caggtggggt gatggccaga ggaatgatgg gcttttgttc 960tgaggggtgt ccgagaggct ggtgtatgca ctgctcacgg accccatgtt ggatctttct 1020ccctttctcc tctccttttt ctcttcacat ctcccccata gcaccctgcc ctcatgggac 1080ctgccctccc tcagccgtca gccatcagcc atggccctcc cagtgcctcc tagccccttc 1140ttccaaggag cagagaggtg gccaccgggg gtggctctgt cctacctcca ctctctgccc 1200ctaaagatgg gaggagacca gcggtccatg ggtctggcct gtgagtctcc ccttgcagcc 1260tggtcactag gcatcacccc cgctttggtt cttcagatgc tcttggggtt cataggggca 1320ggtcctagtc gggcagggcc cctgaccctc ccggcctggc ttcactctcc ctgacggctg 1380ccattggtcc accctttcat agagaggcct gctttgttac aaagctcggg tctccctcct 1440gcagctcggt taagtacccg aggcctctct taagatgtcc agggccccag gcccgcgggc 1500acagccagcc caaaccttgg gccctggaag agtcctccac cccatcacta gagtgctctg 1560accctgggct ttcacgggcc ccattccacc gcctccccaa cttgagcctg tgaccttggg 1620accaaagggg gagtccctcg tctcttgtga ctcagcagag gcagtggcca cgttcaggga 1680ggggccggct ggcctggagg ctcagcccac cctccagctt ttcctcaggg tgtcctgagg 1740tccaagattc tggagcaatc tgacccttct ccaaaggctc tgttatcagc tgggcagtgc 1800cagccaatcc ctggccattt ggccccaggg gacgtgggcc ctg 1843 8 2257 DNA Homosapiens 8 cagggaccta ccccgcgcta cttcacctgg gacgaggtgg cccagcgctcagggtgcgag 60 gagcggtggc tagtgatcga ccgtaaggtg tacaacatca gcgagttcacccgccggcat 120 ccagggggct cccgggtcat cagccactac gccgggcagg atgccacggatccctttgtg 180 gccttccaca tcaacaaggg ccttgtgaag aagtatatga actctctcctgattggagaa 240 ctgtctccag agcagcccag ctttgagccc accaagaata aagagctgacagatgagttc 300 cgggagctgc gggccacagt ggagcggatg gggctcatga aggccaaccatgtcttcttc 360 ctgctgtacc tgctgcacat cttgctgctg gatggtgcag cctggctcaccctttgggtc 420 tttgggacgt cctttttgcc cttcctcctc tgtgcggtgc tgctcagtgcagttcagcag 480 gcccaagctg gatggctgca acatgattat ggccacctgt ctgtctacagaaaacccaag 540 tggaaccacc ttgtccacaa attcgtcatt ggccacttaa agggtgcctctgccaactgg 600 tggaatcatc gccacttcca gcaccacgcc aagcctaaca tcttccacaaggatcccgat 660 gtgaacatgc tgcacgtgtt tgttctgggc gaatggcagc ccatcgagtacggcaagaag 720 aagctgaaat acctgcccta caatcaccag cacgaatact tcttcctgattgggccgccg 780 ctgctcatcc ccatgtattt ccagtaccag atcatcatga ccatgatcgtccataagaac 840 tgggtggacc tggcctgggc cgtcagctac tacatccggt tcttcatcacctacatccct 900 ttctacggca tcctgggagc cctccttttc ctcaacttca tcaggttcctggagagccac 960 tggtttgtgt gggtcacaca gatgaatcac atcgtcatgg agattgaccaggaggcctac 1020 cgtgactggt tcagtagcca gctgacagcc acctgcaacg tggagcagtccttcttcaac 1080 gactggttca gtggacacct taacttccag attgagcacc acctcttccccaccatgccc 1140 cggcacaact tacacaagat cgccccgctg gtgaagtctc tatgtgccaagcatggcatt 1200 gaataccagg agaagccgct actgagggcc ctgctggaca tcatcaggtccctgaagaag 1260 tctgggaagc tgtggctgga cgcctacctt cacaaatgaa gccacagcccccgggacacc 1320 gtggggaagg ggtgcaggtg gggtgatggc cagaggaatg atgggcttttgttctgaggg 1380 gtgtccgaga ggctggtgta tgcactgctc acggacccca tgttggatctttctcccttt 1440 ctcctctcct ttttctcttc acatctcccc catagcaccc tgccctcatgggacctgccc 1500 tccctcagcc gtcagccatc agccatggcc ctcccagtgc ctcctagccccttcttccaa 1560 ggagcagaga ggtggccacc gggggtggct ctgtcctacc tccactctctgcccctaaag 1620 atgggaggag accagcggtc catgggtctg gcctgtgagt ctccccttgcagcctggtca 1680 ctaggcatca cccccgcttt ggttcttcag atgctcttgg ggttcataggggcaggtcct 1740 agtcgggcag ggcccctgac cctcccggcc tggcttcact ctccctgacggctgccattg 1800 gtccaccctt tcatagagag gcctgctttg ttacaaagct cgggtctccctcctgcagct 1860 cggttaagta cccgaggcct ctcttaagat gtccagggcc ccaggcccgcgggcacagcc 1920 agcccaaacc ttgggccctg gaagagtcct ccaccccatc actagagtgctctgaccctg 1980 ggctttcacg ggccccattc caccgcctcc ccaacttgag cctgtgaccttgggaccaaa 2040 gggggagtcc ctcgtctctt gtgactcagc agaggcagtg gccacgttcagggaggggcc 2100 ggctggcctg gaggctcagc ccaccctcca gcttttcctc agggtgtcctgaggtccaag 2160 attctggagc aatctgaccc ttctccaaag gctctgttat cagctgggcagtgccagcca 2220 atccctggcc atttggcccc aggggacgtg ggccctg 2257 9 432 PRTHomo sapiens VARIANT (432)...(432) Xaa = Unknown or other at position432 9 Gln Gly Pro Thr Pro Arg Tyr Phe Thr Trp Asp Glu Val Ala Gln Arg 15 10 15 Ser Gly Cys Glu Glu Arg Trp Leu Val Ile Asp Arg Lys Val Tyr Asn20 25 30 Ile Ser Glu Phe Thr Arg Arg His Pro Gly Gly Ser Arg Val Ile Ser35 40 45 His Tyr Ala Gly Gln Asp Ala Thr Asp Pro Phe Val Ala Phe His Ile50 55 60 Asn Lys Gly Leu Val Lys Lys Tyr Met Asn Ser Leu Leu Ile Gly Glu65 70 75 80 Leu Ser Pro Glu Gln Pro Ser Phe Glu Pro Thr Lys Asn Lys GluLeu 85 90 95 Thr Asp Glu Phe Arg Glu Leu Arg Ala Thr Val Glu Arg Met GlyLeu 100 105 110 Met Lys Ala Asn His Val Phe Phe Leu Leu Tyr Leu Leu HisIle Leu 115 120 125 Leu Leu Asp Gly Ala Ala Trp Leu Thr Leu Trp Val PheGly Thr Ser 130 135 140 Phe Leu Pro Phe Leu Leu Cys Ala Val Leu Leu SerAla Val Gln Ala 145 150 155 160 Gln Ala Gly Trp Leu Gln His Asp Tyr GlyHis Leu Ser Val Tyr Arg 165 170 175 Lys Pro Lys Trp Asn His Leu Val HisLys Phe Val Ile Gly His Leu 180 185 190 Lys Gly Ala Ser Ala Asn Trp TrpAsn His Arg His Phe Gln His His 195 200 205 Ala Lys Pro Asn Ile Phe HisLys Asp Pro Asp Val Asn Met Leu His 210 215 220 Val Phe Val Leu Gly GluTrp Gln Pro Ile Glu Tyr Gly Lys Lys Lys 225 230 235 240 Leu Lys Tyr LeuPro Tyr Asn His Gln His Glu Tyr Phe Phe Leu Ile 245 250 255 Gly Pro ProLeu Leu Ile Pro Met Tyr Phe Gln Tyr Gln Ile Ile Met 260 265 270 Thr MetIle Val His Lys Asn Trp Val Asp Leu Ala Trp Ala Val Ser 275 280 285 TyrTyr Ile Arg Phe Phe Ile Thr Tyr Ile Pro Phe Tyr Gly Ile Leu 290 295 300Gly Ala Leu Leu Phe Leu Asn Phe Ile Arg Phe Leu Glu Ser His Trp 305 310315 320 Phe Val Trp Val Thr Gln Met Asn His Ile Val Met Glu Ile Asp Gln325 330 335 Glu Ala Tyr Arg Asp Trp Phe Ser Ser Gln Leu Thr Ala Thr CysAsn 340 345 350 Val Glu Gln Ser Phe Phe Asn Asp Trp Phe Ser Gly His LeuAsn Phe 355 360 365 Gln Ile Glu His His Leu Phe Pro Thr Met Pro Arg HisAsn Leu His 370 375 380 Lys Ile Ala Pro Leu Val Lys Ser Leu Cys Ala LysHis Gly Ile Glu 385 390 395 400 Tyr Gln Glu Lys Pro Leu Leu Arg Ala LeuLeu Asp Ile Ile Arg Ser 405 410 415 Leu Lys Lys Ser Gly Lys Leu Trp LeuAsp Ala Tyr Leu His Lys Xaa 420 425 430 10 447 PRT Homo sapiens VARIANT(447)...(447) Xaa = Unknown or Other at position 447 10 Met Gly Thr AspGln Gly Lys Thr Phe Thr Trp Glu Glu Leu Ala Ala 1 5 10 15 His Asn ThrLys Asp Asp Leu Leu Leu Ala Ile Arg Gly Arg Val Tyr 20 25 30 Asp Val ThrLys Phe Leu Ser Arg His Pro Gly Gly Val Asp Thr Leu 35 40 45 Leu Leu GlyAla Gly Arg Asp Val Thr Pro Val Phe Glu Met Tyr His 50 55 60 Ala Phe GlyAla Ala Asp Ala Ile Met Lys Lys Tyr Tyr Val Gly Thr 65 70 75 80 Leu ValSer Asn Glu Leu Pro Ile Phe Pro Glu Pro Thr Val Phe His 85 90 95 Lys ThrIle Lys Thr Arg Val Glu Gly Tyr Phe Thr Asp Arg Asn Ile 100 105 110 AspPro Lys Asn Arg Pro Glu Ile Trp Gly Arg Tyr Ala Leu Ile Phe 115 120 125Gly Ser Leu Ile Ala Ser Tyr Tyr Ala Gln Leu Phe Val Pro Phe Val 130 135140 Val Glu Arg Thr Trp Leu Gln Val Val Phe Ala Ile Ile Met Gly Phe 145150 155 160 Ala Cys Ala Gln Val Gly Leu Asn Pro Leu His Asp Ala Ser HisPhe 165 170 175 Ser Val Thr His Asn Pro Thr Val Trp Lys Ile Leu Gly AlaThr His 180 185 190 Asp Phe Phe Asn Gly Ala Ser Tyr Leu Val Trp Met TyrGln His Met 195 200 205 Leu Gly His His Pro Tyr Thr Asn Ile Ala Gly AlaAsp Pro Asp Val 210 215 220 Ser Thr Ser Glu Pro Asp Val Arg Arg Ile LysPro Asn Gln Lys Trp 225 230 235 240 Phe Val Asn His Ile Asn Gln His MetPhe Val Pro Phe Leu Tyr Gly 245 250 255 Leu Leu Ala Phe Lys Val Arg IleGln Asp Ile Asn Ile Leu Tyr Phe 260 265 270 Val Lys Thr Asn Asp Ala IleArg Val Asn Pro Ile Ser Thr Trp His 275 280 285 Thr Val Met Phe Trp GlyGly Lys Ala Phe Phe Val Trp Tyr Arg Leu 290 295 300 Ile Val Pro Leu GlnTyr Leu Pro Leu Gly Lys Val Leu Leu Leu Phe 305 310 315 320 Thr Val AlaAsp Met Val Ser Ser Tyr Trp Leu Ala Leu Thr Phe Gln 325 330 335 Ala AsnHis Val Val Glu Glu Val Gln Trp Pro Leu Pro Asp Glu Asn 340 345 350 GlyIle Ile Gln Lys Asp Trp Ala Ala Met Gln Val Glu Thr Thr Gln 355 360 365Asp Tyr Ala His Asp Ser His Leu Trp Thr Ser Ile Thr Gly Ser Leu 370 375380 Asn Tyr Gln Ala Val His His Leu Phe Pro Asn Val Ser Gln His His 385390 395 400 Tyr Pro Asp Ile Leu Ala Ile Ile Lys Asn Thr Cys Ser Glu TyrLys 405 410 415 Val Pro Tyr Leu Val Lys Asp Thr Phe Trp Gln Ala Phe AlaSer His 420 425 430 Leu Glu His Leu Arg Val Leu Gly Leu Arg Pro Lys GluGlu Xaa 435 440 445 11 458 PRT Homo sapiens VARIANT (458)...(458) Xaa =Unknown or other at position 458 11 Met Ala Ala Ala Pro Ser Val Arg ThrPhe Thr Arg Ala Glu Val Leu 1 5 10 15 Asn Ala Glu Ala Leu Asn Glu GlyLys Lys Asp Ala Glu Ala Pro Phe 20 25 30 Leu Met Ile Ile Asp Asn Lys ValTyr Asp Val Arg Glu Phe Val Pro 35 40 45 Asp His Pro Gly Gly Ser Val IleLeu Thr His Val Gly Lys Asp Gly 50 55 60 Thr Asp Val Phe Asp Thr Phe HisPro Glu Ala Ala Trp Glu Thr Leu 65 70 75 80 Ala Asn Phe Tyr Val Gly AspIle Asp Glu Ser Asp Arg Asp Ile Lys 85 90 95 Asn Asp Asp Phe Ala Ala GluVal Arg Lys Leu Arg Thr Leu Phe Gln 100 105 110 Ser Leu Gly Tyr Tyr AspSer Ser Lys Ala Tyr Tyr Ala Phe Lys Val 115 120 125 Ser Phe Asn Leu CysIle Trp Gly Leu Ser Thr Val Ile Val Ala Lys 130 135 140 Trp Gly Gln ThrSer Thr Leu Ala Asn Val Leu Ser Ala Ala Leu Leu 145 150 155 160 Gly LeuPhe Trp Gln Gln Cys Gly Trp Leu Ala His Asp Phe Leu His 165 170 175 HisGln Val Phe Gln Asp Arg Phe Trp Gly Asp Leu Phe Gly Ala Phe 180 185 190Leu Gly Gly Val Cys Gln Gly Phe Ser Ser Ser Trp Trp Lys Asp Lys 195 200205 His Asn Thr His His Ala Ala Pro Asn Val His Gly Glu Asp Pro Asp 210215 220 Ile Asp Thr His Pro Leu Leu Thr Trp Ser Glu His Ala Leu Glu Met225 230 235 240 Phe Ser Asp Val Pro Asp Glu Glu Leu Thr Arg Met Trp SerArg Phe 245 250 255 Met Val Leu Asn Gln Thr Trp Phe Tyr Phe Pro Ile LeuSer Phe Ala 260 265 270 Arg Leu Ser Trp Cys Leu Gln Ser Ile Leu Phe ValLeu Pro Asn Gly 275 280 285 Gln Ala His Lys Pro Ser Gly Ala Arg Val ProIle Ser Leu Val Glu 290 295 300 Gln Leu Ser Leu Ala Met His Trp Thr TrpTyr Leu Ala Thr Met Phe 305 310 315 320 Leu Phe Ile Lys Asp Pro Val AsnMet Leu Val Tyr Phe Leu Val Ser 325 330 335 Gln Ala Val Cys Gly Asn LeuLeu Ala Ile Val Phe Ser Leu Asn His 340 345 350 Asn Gly Met Pro Val IleSer Lys Glu Glu Ala Val Asp Met Asp Phe 355 360 365 Phe Thr Lys Gln IleIle Thr Gly Arg Asp Val His Pro Gly Leu Phe 370 375 380 Ala Asn Trp PheThr Gly Gly Leu Asn Tyr Gln Ile Glu His His Leu 385 390 395 400 Phe ProSer Met Pro Arg His Asn Phe Ser Lys Ile Gln Pro Ala Val 405 410 415 GluThr Leu Cys Lys Lys Tyr Asn Val Arg Tyr His Thr Thr Gly Met 420 425 430Ile Glu Gly Thr Ala Glu Val Phe Ser Arg Leu Asn Glu Val Ser Lys 435 440445 Ala Ala Ser Lys Met Gly Lys Ala Gln Xaa 450 455 12 444 PRT Homosapiens 12 Met Ala Pro Asp Pro Val Ala Ala Glu Thr Ala Ala Gln Gly ProThr 1 5 10 15 Pro Arg Tyr Phe Thr Trp Asp Glu Val Ala Gln Arg Ser GlyCys Glu 20 25 30 Glu Arg Trp Leu Val Ile Asp Arg Lys Val Tyr Asn Ile SerGlu Phe 35 40 45 Thr Arg Arg His Pro Gly Gly Ser Arg Val Ile Ser His TyrAla Gly 50 55 60 Gln Asp Ala Thr Asp Pro Phe Val Ala Phe His Ile Asn LysGly Leu 65 70 75 80 Val Lys Lys Tyr Met Asn Ser Leu Leu Ile Gly Glu LeuSer Pro Glu 85 90 95 Gln Pro Ser Phe Glu Pro Thr Lys Asn Lys Glu Leu ThrAsp Glu Phe 100 105 110 Arg Glu Leu Arg Ala Thr Val Glu Arg Met Gly LeuMet Lys Ala Asn 115 120 125 His Val Phe Phe Leu Leu Tyr Leu Leu His IleLeu Leu Leu Asp Gly 130 135 140 Ala Ala Trp Leu Thr Leu Trp Val Phe GlyThr Ser Phe Leu Pro Phe 145 150 155 160 Leu Leu Cys Ala Val Leu Leu SerAla Val Gln Ala Gln Ala Gly Trp 165 170 175 Leu Gln His Asp Phe Gly HisLeu Ser Val Phe Ser Thr Ser Lys Trp 180 185 190 Asn His Leu Leu His HisPhe Val Ile Gly His Leu Lys Gly Ala Pro 195 200 205 Ala Ser Trp Trp AsnHis Met His Phe Gln His His Ala Lys Pro Asn 210 215 220 Cys Phe Arg LysAsp Pro Asp Ile Asn Met His Pro Phe Phe Phe Ala 225 230 235 240 Leu GlyLys Ile Leu Ser Val Glu Leu Gly Lys Gln Lys Lys Lys Tyr 245 250 255 MetPro Tyr Asn His Gln His Lys Tyr Phe Phe Leu Ile Gly Pro Pro 260 265 270Ala Leu Leu Pro Leu Tyr Phe Gln Trp Tyr Ile Phe Tyr Phe Val Ile 275 280285 Gln Arg Lys Lys Trp Val Asp Leu Ala Trp Met Ile Thr Phe Tyr Val 290295 300 Arg Phe Phe Leu Thr Tyr Val Pro Leu Leu Gly Leu Lys Ala Phe Leu305 310 315 320 Gly Leu Phe Phe Ile Val Arg Phe Leu Glu Ser Asn Trp PheVal Trp 325 330 335 Val Thr Gln Met Asn His Ile Pro Met His Ile Asp HisAsp Arg Asn 340 345 350 Met Asp Trp Val Ser Thr Gln Leu Leu Ala Thr CysAsn Val His Lys 355 360 365 Ser Ala Phe Asn Asp Trp Phe Ser Gly His LeuAsn Phe Gln Ile Glu 370 375 380 His His Leu Phe Pro Thr Met Pro Arg HisAsn Tyr His Lys Val Ala 385 390 395 400 Pro Leu Val Gln Ser Leu Cys AlaLys Arg Gly Ile Glu Tyr Gln Ser 405 410 415 Lys Pro Leu Leu Ser Ala PheAla Asp Ile Ile His Ser Leu Lys Glu 420 425 430 Ser Gly Gln Leu Trp LeuAsp Ala Tyr Leu His Gln 435 440 13 864 DNA Homo sapiens 13 ctcctggagcccgtcagtat cggcggaatt ccggcagttc aggcccaggc tggctggctg 60 cagcatgactttgggcacct gtcggtcttc agcacctcaa agtggaacca tctgctacat 120 cattttgtgattggccacct gaagggggcc cccgccagtt ggtggaacca catgcacttc 180 cagcaccatgccaagcccaa ctgcttccgc aaagacccag acatcaacat gcatcccttc 240 ttctttgccttggggaagat cctctctgtg gagcttggga aacagaagaa aaaatatatg 300 ccgtacaaccaccagcacaa atacttcttc ctaattgggc ccccagcctt gctgcctctc 360 tacttccagtggtatatttt ctattttgtt atccagcgaa agaagtgggt ggacttggcc 420 tggatgattaccttctacgt ccgcttcttc ctcacttatg tgccactatt ggggctgaaa 480 gccttcctgggccttttctt catagtcagg ttcctggaaa gcaactggtt tgtgtgggtg 540 acacagatgaaccatattcc catgcacatt gatcatgacc ggaacatgga ctgggtttcc 600 acccagctccaggccacatg caatgtccac aagtctgcct tcaatgactg gttcagtgga 660 cacctcaacttccagattga gcaccatctt tttcccacga tgcctcgaca caattaccac 720 aaagtggctcccctggtgca gtccttgtgt gccaagcatg gcatagagta ccagtccaag 780 cccctgctgtcagccttcgc cgacatcatc cactcactaa aggagtcagg gcagctctgg 840 ctagatgcctatcttcacca ataa 864 14 287 PRT Homo sapiens 14 Leu Leu Glu Pro Val SerIle Gly Gly Ile Pro Ala Val Gln Ala Gln 1 5 10 15 Ala Gly Trp Leu GlnHis Asp Phe Gly His Leu Ser Val Phe Ser Thr 20 25 30 Ser Lys Trp Asn HisLeu Leu His His Phe Val Ile Gly His Leu Lys 35 40 45 Gly Ala Pro Ala SerTrp Trp Asn His Met His Phe Gln His His Ala 50 55 60 Lys Pro Asn Cys PheArg Lys Asp Pro Asp Ile Asn Met His Pro Phe 65 70 75 80 Phe Phe Ala LeuGly Lys Ile Leu Ser Val Glu Leu Gly Lys Gln Lys 85 90 95 Lys Lys Tyr MetPro Tyr Asn His Gln His Lys Tyr Phe Phe Leu Ile 100 105 110 Gly Pro ProAla Leu Leu Pro Leu Tyr Phe Gln Trp Tyr Ile Phe Tyr 115 120 125 Phe ValIle Gln Arg Lys Lys Trp Val Asp Leu Ala Trp Met Ile Thr 130 135 140 PheTyr Val Arg Phe Phe Leu Thr Tyr Val Pro Leu Leu Gly Leu Lys 145 150 155160 Ala Phe Leu Gly Leu Phe Phe Ile Val Arg Phe Leu Glu Ser Asn Trp 165170 175 Phe Val Trp Val Thr Gln Met Asn His Ile Pro Met His Ile Asp His180 185 190 Asp Arg Asn Met Asp Trp Val Ser Thr Gln Leu Gln Ala Thr CysAsn 195 200 205 Val His Lys Ser Ala Phe Asn Asp Trp Phe Ser Gly His LeuAsn Phe 210 215 220 Gln Ile Glu His His Leu Phe Pro Thr Met Pro Arg HisAsn Tyr His 225 230 235 240 Lys Val Ala Pro Leu Val Gln Ser Leu Cys AlaLys His Gly Ile Glu 245 250 255 Tyr Gln Ser Lys Pro Leu Leu Ser Ala PheAla Asp Ile Ile His Ser 260 265 270 Leu Lys Glu Ser Gly Gln Leu Trp LeuAsp Ala Tyr Leu His Gln 275 280 285 15 288 PRT Homo sapiens VARIANT(288)...(288) Xaa = Unknown or other at position 288 15 Leu Leu Glu ProVal Ser Ile Gly Gly Ile Pro Ala Val Gln Ala Gln 1 5 10 15 Ala Gly TrpLeu Gln His Asp Phe Gly His Leu Ser Val Phe Ser Thr 20 25 30 Ser Lys TrpAsn His Leu Leu His His Phe Val Ile Gly His Leu Lys 35 40 45 Gly Ala ProAla Ser Trp Trp Asn His Met His Phe Gln His His Ala 50 55 60 Lys Pro AsnCys Phe Arg Lys Asp Pro Asp Ile Asn Met His Pro Phe 65 70 75 80 Phe PheAla Leu Gly Lys Ile Leu Ser Val Glu Leu Gly Lys Gln Lys 85 90 95 Lys LysTyr Met Pro Tyr Asn His Gln His Lys Tyr Phe Phe Leu Ile 100 105 110 GlyPro Pro Ala Leu Leu Pro Leu Tyr Phe Gln Trp Tyr Ile Phe Tyr 115 120 125Phe Val Ile Gln Arg Lys Lys Trp Val Asp Leu Ala Trp Met Ile Thr 130 135140 Phe Tyr Val Arg Phe Phe Leu Thr Tyr Val Pro Leu Leu Gly Leu Lys 145150 155 160 Ala Phe Leu Gly Leu Phe Phe Ile Val Arg Phe Leu Glu Ser AsnTrp 165 170 175 Phe Val Trp Val Thr Gln Met Asn His Ile Pro Met His IleAsp His 180 185 190 Asp Arg Asn Met Asp Trp Val Ser Thr Gln Leu Gln AlaThr Cys Asn 195 200 205 Val His Lys Ser Ala Phe Asn Asp Trp Phe Ser GlyHis Leu Asn Phe 210 215 220 Gln Ile Glu His His Leu Phe Pro Thr Met ProArg His Asn Tyr His 225 230 235 240 Lys Val Ala Pro Leu Val Gln Ser LeuCys Ala Lys His Gly Ile Glu 245 250 255 Tyr Gln Ser Lys Pro Leu Leu SerAla Phe Ala Asp Ile Ile His Ser 260 265 270 Leu Lys Glu Ser Gly Gln LeuTrp Leu Asp Ala Tyr Leu His Gln Xaa 275 280 285 16 315 PRT Homo sapiensVARIANT (315)...(315) Xaa = Unknown or other at position 315 16 Ala SerTyr Tyr Ala Gln Leu Phe Val Pro Phe Val Val Glu Arg Thr 1 5 10 15 TrpLeu Gln Val Val Phe Ala Ile Ile Met Gly Phe Ala Cys Ala Gln 20 25 30 ValGly Leu Asn Pro Leu His Asp Ala Ser His Phe Ser Val Thr His 35 40 45 AsnPro Thr Val Trp Lys Ile Leu Gly Ala Thr His Asp Phe Phe Asn 50 55 60 GlyAla Ser Tyr Leu Val Trp Met Tyr Gln His Met Leu Gly His His 65 70 75 80Pro Tyr Thr Asn Ile Ala Gly Ala Asp Pro Asp Val Ser Thr Ser Glu 85 90 95Pro Asp Val Arg Arg Ile Lys Pro Asn Gln Lys Trp Phe Val Asn His 100 105110 Ile Asn Gln His Met Phe Val Pro Phe Leu Tyr Gly Leu Leu Ala Phe 115120 125 Lys Val Arg Ile Gln Asp Ile Asn Ile Leu Tyr Phe Val Lys Thr Asn130 135 140 Asp Ala Ile Arg Val Asn Pro Ile Ser Thr Trp His Thr Val MetPhe 145 150 155 160 Trp Gly Gly Lys Ala Phe Phe Val Trp Tyr Arg Leu IleVal Pro Leu 165 170 175 Gln Tyr Leu Pro Leu Gly Lys Val Leu Leu Leu PheThr Val Ala Asp 180 185 190 Met Val Ser Ser Tyr Trp Leu Ala Leu Thr PheGln Ala Asn His Val 195 200 205 Val Glu Glu Val Gln Trp Pro Leu Pro AspGlu Asn Gly Ile Ile Gln 210 215 220 Lys Asp Trp Ala Ala Met Gln Val GluThr Thr Gln Asp Tyr Ala His 225 230 235 240 Asp Ser His Leu Trp Thr SerIle Thr Gly Ser Leu Asn Tyr Gln Ala 245 250 255 Val His His Leu Phe ProAsn Val Ser Gln His His Tyr Pro Asp Ile 260 265 270 Leu Ala Ile Ile LysAsn Thr Cys Ser Glu Tyr Lys Val Pro Tyr Leu 275 280 285 Val Lys Asp ThrPhe Trp Gln Ala Phe Ala Ser His Leu Glu His Leu 290 295 300 Arg Val LeuGly Leu Arg Pro Lys Glu Glu Xaa 305 310 315 17 323 PRT Homo sapiensVARIANT (323)...(323) Xaa = Unknown or other at position 323 17 Gly LeuSer Thr Val Ile Val Ala Lys Trp Gly Gln Thr Ser Thr Leu 1 5 10 15 AlaAsn Val Leu Ser Ala Ala Leu Leu Gly Leu Phe Trp Gln Gln Cys 20 25 30 GlyTrp Leu Ala His Asp Phe Leu His His Gln Val Phe Gln Asp Arg 35 40 45 PheTrp Gly Asp Leu Phe Gly Ala Phe Leu Gly Gly Val Cys Gln Gly 50 55 60 PheSer Ser Ser Trp Trp Lys Asp Lys His Asn Thr His His Ala Ala 65 70 75 80Pro Asn Val His Gly Glu Asp Pro Asp Ile Asp Thr His Pro Leu Leu 85 90 95Thr Trp Ser Glu His Ala Leu Glu Met Phe Ser Asp Val Pro Asp Glu 100 105110 Glu Leu Thr Arg Met Trp Ser Arg Phe Met Val Leu Asn Gln Thr Trp 115120 125 Phe Tyr Phe Pro Ile Leu Ser Phe Ala Arg Leu Ser Trp Cys Leu Gln130 135 140 Ser Ile Leu Phe Val Leu Pro Asn Gly Gln Ala His Lys Pro SerGly 145 150 155 160 Ala Arg Val Pro Ile Ser Leu Val Glu Gln Leu Ser LeuAla Met His 165 170 175 Trp Thr Trp Tyr Leu Ala Thr Met Phe Leu Phe IleLys Asp Pro Val 180 185 190 Asn Met Leu Val Tyr Phe Leu Val Ser Gln AlaVal Cys Gly Asn Leu 195 200 205 Leu Ala Ile Val Phe Ser Leu Asn His AsnGly Met Pro Val Ile Ser 210 215 220 Lys Glu Glu Ala Val Asp Met Asp PhePhe Thr Lys Gln Ile Ile Thr 225 230 235 240 Gly Arg Asp Val His Pro GlyLeu Phe Ala Asn Trp Phe Thr Gly Gly 245 250 255 Leu Asn Tyr Gln Ile GluHis His Leu Phe Pro Ser Met Pro Arg His 260 265 270 Asn Phe Ser Lys IleGln Pro Ala Val Glu Thr Leu Cys Lys Lys Tyr 275 280 285 Asn Val Arg TyrHis Thr Thr Gly Met Ile Glu Gly Thr Ala Glu Val 290 295 300 Phe Ser ArgLeu Asn Glu Val Ser Lys Ala Ala Ser Lys Met Gly Lys 305 310 315 320 AlaGln Xaa 18 356 PRT Homo sapiens VARIANT (294)...(294) Xaa = Unknown orother at position 294 18 Val Phe Tyr Phe Gly Asn Gly Trp Ile Pro Thr LeuIle Thr Ala Phe 1 5 10 15 Val Leu Ala Thr Ser Gln Ala Gln Ala Gly TrpLeu Gln His Asp Tyr 20 25 30 Gly His Leu Ser Val Tyr Arg Lys Pro Lys TrpAsn His Leu Val His 35 40 45 Lys Phe Val Ile Gly His Leu Lys Gly Ala SerAla Asn Trp Trp Asn 50 55 60 His Arg His Phe Gln His His Ala Lys Pro AsnIle Phe His Lys Asp 65 70 75 80 Pro Asp Val Asn Met Leu His Val Phe ValLeu Gly Glu Trp Gln Pro 85 90 95 Ile Glu Tyr Gly Lys Lys Lys Leu Lys TyrLeu Pro Tyr Asn His Gln 100 105 110 His Glu Tyr Phe Phe Leu Ile Gly ProPro Leu Leu Ile Pro Met Tyr 115 120 125 Phe Gln Tyr Gln Ile Ile Met ThrMet Ile Val His Lys Asn Trp Val 130 135 140 Asp Leu Ala Trp Ala Val SerTyr Tyr Ile Arg Phe Phe Ile Thr Ile 145 150 155 160 Pro Phe Tyr Gly IleLeu Gly Ala Leu Leu Phe Leu Asn Phe Ile Arg 165 170 175 Phe Leu Glu SerHis Trp Phe Val Trp Val Thr Gln Met Asn His Ile 180 185 190 Val Met GluIle Asp Gln Glu Ala Tyr Arg Asp Trp Phe Ser Ser Gln 195 200 205 Leu ThrAla Thr Cys Asn Val Glu Gln Ser Phe Phe Asn Asp Trp Phe 210 215 220 SerGly His Leu Asn Phe Gln Ile Glu His His Leu Phe Pro Thr Met 225 230 235240 Pro Arg His Asn Leu His Lys Ile Ala Pro Leu Val Lys Ser Leu Cys 245250 255 Ala Lys His Gly Ile Glu Tyr Gln Glu Lys Pro Leu Leu Arg Ala Leu260 265 270 Leu Asp Ile Ile Arg Ser Leu Lys Lys Ser Gly Lys Leu Trp LeuAsp 275 280 285 Ala Tyr Leu His Lys Xaa Ser His Ser Pro Arg Asp Thr ValGly Lys 290 295 300 Gly Cys Arg Trp Gly Asp Gly Gln Arg Asn Asp Gly LeuLeu Phe Xaa 305 310 315 320 Gly Val Ser Glu Arg Leu Val Tyr Ala Leu LeuThr Asp Pro Met Leu 325 330 335 Asp Leu Ser Pro Phe Leu Leu Ser Phe PheSer Ser His Leu Pro His 340 345 350 Ser Thr Leu Pro 355 19 219 PRT Homosapiens 19 Leu Leu Glu Pro Val Ser Ile Gly Gly Ile Pro Ala Val Gln AlaGln 1 5 10 15 Ala Gly Trp Leu Gln His Asp Phe Gly His Leu Ser Val PheSer Thr 20 25 30 Ser Lys Trp Asn His Leu Leu His His Phe Val Ile Gly HisLeu Lys 35 40 45 Gly Ala Pro Ala Ser Trp Trp Asn His Met His Phe Gln HisHis Ala 50 55 60 Lys Pro Asn Cys Phe Arg Lys Asp Pro Asp Ile Asn Met HisPro Phe 65 70 75 80 Phe Phe Ala Leu Gly Lys Ile Leu Ser Val Glu Leu GlyLys Gln Lys 85 90 95 Lys Lys Tyr Met Pro Tyr Asn His Gln His Lys Tyr PhePhe Leu Ile 100 105 110 Gly Pro Pro Ala Leu Leu Pro Leu Tyr Phe Gln TrpTyr Ile Phe Tyr 115 120 125 Phe Val Ile Gln Arg Lys Lys Trp Val Asp LeuAla Trp Met Ile Thr 130 135 140 Phe Tyr Val Arg Phe Phe Leu Thr Tyr ValPro Leu Leu Gly Leu Lys 145 150 155 160 Ala Phe Leu Gly Leu Phe Phe IleVal Arg Phe Leu Glu Ser Asn Trp 165 170 175 Phe Val Trp Val Thr Gln MetAsn His Ile Pro Met His Ile Asp His 180 185 190 Asp Arg Asn Met Asp TrpVal Ser Thr Gln Leu Gln Ala Thr Cys Asn 195 200 205 Val His Lys Ser AlaPhe Asn Asp Trp Phe Ser 210 215 20 182 PRT Homo sapiens VARIANT(128)...(128) Xaa = Unknown or other at position 128 20 Leu His Ile LeuLeu Leu Asp Gly Ala Ala Trp Leu Thr Leu Trp Val 1 5 10 15 Phe Gly ThrSer Phe Leu Pro Phe Leu Leu Cys Ala Val Leu Leu Ser 20 25 30 Ala Val GlnAla Gln Ala Gly Trp Leu Gln His Asp Phe Gly His Leu 35 40 45 Ser Val PheSer Thr Ser Lys Trp Asn His Leu Leu His His Phe Val 50 55 60 Ile Gly HisLeu Lys Gly Ala Pro Ala Ser Trp Trp Asn His Met His 65 70 75 80 Phe GlnHis His Ala Lys Pro Asn Cys Phe Arg Lys Asp Pro Asp Ile 85 90 95 Asn MetHis Pro Phe Phe Phe Ala Leu Gly Lys Ile Leu Ser Val Glu 100 105 110 LeuGly Lys Gln Lys Lys Lys Tyr Met Pro Tyr Asn His Gln His Xaa 115 120 125Tyr Phe Phe Leu Ile Gly Pro Pro Ala Leu Leu Pro Leu Tyr Phe Gln 130 135140 Trp Tyr Ile Phe Tyr Phe Val Ile Gln Arg Lys Lys Trp Val Asp Leu 145150 155 160 Ala Trp Ile Ser Lys Gln Glu Tyr Asp Glu Ala Gly Leu Pro LeuSer 165 170 175 Thr Ala Asn Ala Ser Lys 180 21 179 PRT Homo sapiensVARIANT (1)...(1) Xaa = Unknown or other at position 1 21 Xaa Leu AspLeu Pro Thr Asn Met Met Glu Xaa Arg Lys Ala Ala Ala 1 5 10 15 Glu LeuXaa Ala Ala Glu Thr Ala Ala Gln Gly Pro Thr Pro Arg Tyr 20 25 30 Phe ThrTrp Asp Glu Val Ala Gln Arg Ser Gly Cys Glu Glu Arg Trp 35 40 45 Leu ValIle Asp Arg Lys Val Tyr Asn Ile Ser Glu Phe Thr Arg Arg 50 55 60 His ProGly Gly Ser Arg Val Ile Ser His Tyr Ala Gly Gln Asp Ala 65 70 75 80 ThrAsp Pro Phe Val Ala Phe His Ile Asn Lys Gly Leu Val Lys Lys 85 90 95 TyrMet Asn Ser Leu Leu Ile Gly Glu Leu Ser Pro Glu Gln Pro Ser 100 105 110Phe Glu Pro Thr Lys Asn Lys Glu Leu Thr Asp Glu Phe Arg Glu Leu 115 120125 Arg Ala Thr Val Glu Gln Arg Phe Pro Val Xaa Phe Leu Thr Cys Thr 130135 140 Gly Ala His Gly Phe Phe Ser Leu Glu Val Pro Gly Leu Pro Asp Ser145 150 155 160 Asn Lys Xaa Phe Ser Trp Thr Ser Arg Pro Ile Xaa Trp AsnLys Gly 165 170 175 Lys Arg Pro 22 133 PRT Homo sapiens 22 Ala Glu GlnSer Asp Glu Ala Val Lys Tyr Tyr Thr Leu Glu Glu Ile 1 5 10 15 Gln LysHis Asn His Ser Lys Ser Thr Trp Leu Ile Leu His His Lys 20 25 30 Val TyrAsp Leu Thr Lys Phe Leu Glu Glu His Pro Gly Gly Glu Glu 35 40 45 Val LeuArg Glu Gln Ala Gly Gly Asp Ala Thr Glu Asn Phe Glu Asp 50 55 60 Val GlyHis Ser Thr Asp Ala Arg Glu Met Ser Lys Thr Phe Ile Ile 65 70 75 80 GlyGlu Leu His Pro Asp Asp Arg Pro Lys Leu Asn Lys Pro Pro Glu 85 90 95 ThrLeu Ile Thr Thr Ile Asp Ser Ser Ser Ser Trp Trp Thr Asn Trp 100 105 110Val Ile Pro Ala Ile Ser Ala Val Ala Val Ala Leu Met Tyr Arg Leu 115 120125 Tyr Met Ala Glu Asp 130 23 300 DNA Homo sapiens 23 cccgaccaatatgatggaat aaaggaaagc ggccgctgaa ttataggccg ccgagaccgc 60 ggctcagggacctaccccgc gttacttcac atgggacgag gtggcccagc gctcagggtg 120 cgaggagcggtggcttgtga tcgaccgtaa ggtgtacaac atcagcgagt tcacccgccg 180 gcatccagggggctcccggg tcatcagcca ctacgccggg caggatgcca cggatccctt 240 cgtggccttccacatcaaca agggccttgt gaagaagtat atgaactctc tcctgattgg 300 24 300 DNAHomo sapiens 24 cccggcgcgc ggcgtcgcca ggccagctat ggcccccgac ccggtggccgccgagaccgc 60 ggctcaggga cctaccccgc gctacttcac ctgggacgag gtggcccagcgctcagggtg 120 cgaggagcgg tggctagtga tcgaccgtaa ggtgtacaac atcagcgagttcacccgccg 180 gcatccaggg ggctcccggg tcatcagcca ctacgccggg caggatgccacggtgagcgc 240 agccaggcgg gggcacagga gagggcggga ccggaggctg agtgcaggggagacagagtt 300 25 360 DNA Homo sapiens 25 aatacgactc actatagggctcgagcggcc gcccgggcag gtccggacct gccaacgtga 60 atcttatcgc catggaccttaccttgcaca acccaaagta gctgccttgg ggcagggggt 120 ggccagagtg cttagggaaatgtggagccc tacccagaac aacggtggag ggaaagggaa 180 gaaacgcaga agtgccccagttcggacgta gggaagtctt cctcttcgtg gtttttggag 240 aaccctagct aagagaggaaagggacttat tgaaagaccc gcaagaaggg acggaagtct 300 catagccctg agaggatccctttgtggcct tccacatcaa caagggcctt gtgaagaagt 360 26 419 DNA Homo sapiens26 ccccgcccca cacgccgcat cacttacagg gcccggggct gccggacctg ccaacgtgaa 60tcttatcgcc atggacctta ccttgcacaa cccaaagtag ctgccttggg gcagggggtg 120gccagagtgc ttagggaaat gtggagccct acccagaaca acggtggagg gaaagggaag 180aaacgcagaa gtgccccagt tcggacgtag ggaagtcttc ctcttcgtgg tttttggaga 240accctagcta agagaggaaa gggacttatt gaaagacccg caagaaggga cggaagtcta 300accctagcta agagaggaaa gggacttatt gaaagacccg caagaaggga cggaagtctc 360atagccctga gaggtgaagc cagctggagt tgatgggtcg aatggggacc tagagaact 419 27240 DNA Homo sapiens 27 tatagggctc gagcggccgc ccgggcaggt gcccggaggcgcctgatcat acctgttgcc 60 cggtgattgg gtgtcctgcg gatgcgggat gaaaaggcgggagagaggcc tggaaaagtg 120 gagtctgggg agtggggatg gaggccaaca acacgcacacacaaacaaag ggtcccgcct 180 ccctgccgtg cattccatct gcagccccga gcctcaggatccctttgtgg ccttccacat 240 28 239 DNA Homo sapiens 28 cgagccaaacaccgactaat tcggaggaaa gcccggaggc gcctgatcat acctgttgcc 60 cggtgattgggtgtcctgcg gatgcgggat gaaaaggcgg gagagaggcc tggagaagtg 120 gagtctggggagtggggatg gaggccaaca acacgcacac acaaacaaag ggtcccgcct 180 ccctgccgtgcattccatct gcagccccga gcctcaggtc tctgggcggg gacagaacc 239 29 300 DNAHomo sapiens 29 cgagcggccg cccgggcagg tctagaattc agcggccgct gaagccgcgtctggacctag 60 gtgccggtct ccactcgcca gcaggagcgg agagggagca ggaaaggagcccattctcga 120 ggatggggct gaaacgggaa gcttggggag accgctgcct tggggacccctgcgtcgtgt 180 gaagactgga ggacgcggaa gggacagcgc tggccgggga gggcaagcggccgctggcga 240 tccctttgtg gccttccaca tcaacaaggg ccttgtgaag aagtatatgaactctctcct 300 30 299 DNA Homo sapiens 30 agggagtcac atcctgtctcgatggctagg agaggcagcg cagccgcgtc tggacctagg 60 tgccggtctc cactcgccagcaggagcgga gagggagcag gaaaggagcc cattctcgag 120 gatggggctg aaacgggaagcttggggaga ccgctgcctt ggggacccct gcgtcgtgtg 180 aagactggag gacgcggaagggacagcgct ggccggggag ggcaagcggc cgctggcgta 240 cataagggat tgggaatggcatacacttag cgaggacccc cagagctgtt ctcgaatcg 299 31 286 DNA Homo sapiens31 ttattccctt atttgtccct gcccatgtcc tgctgattgg tccattttac ctctagctag 60ctaaagagca cggattggtg cattttgcaa acctctggct acagaggggt tctccaggtc 120tgcactcgac ccaggaagtc catctggctt cacctctcac ttcaacttgg gtacagcctt 180ctggcgggca ggaagatggc ctttggtgcg aacactgccg gagtccaggg ggctggctcc 240ctcacctttc atcttctccc ggcacttgca ggatcccttt gtggcc 286 32 286 DNA Homosapiens 32 atagagcact gattggtcca ttttacaggg tgctgattgg tccattttacctctagctag 60 ctaaagagca cggattggtg cattttacaa acctctagct acagaaaagttctccaagtc 120 tgcactcgac ccaggaagtc catctggctt cacctctcac ttcaacttgggtacagcctt 180 ctggcgggca ggaggatggc ctttggtgcg aacactgccg gagtccagggggctggctcc 240 ctcacctttc atcttctccc ggcacttgca ggatcccttt gtggcc 286 334698 DNA Homo sapiens 33 actagaaccg ctgttcctac cgcggcgccc cctgggagccaacgccgcga tgcccgcctg 60 acgtcaggaa gtcgaatccg gcggcgacgc ttttagggagcccgcgaggg ggcgcgtgtt 120 ggcagcccag ctgtgagttg cccaagaccc accgggggacgggatctcgc tccccgcgcc 180 acgaggctcg gccaatggga acgcgcgctg cgaggcccgccggtctgccc tgcggtgctg 240 aaaacccggc gcgcaggcgg ctggctctgg gcgcgcgccagcaaatccac tcctggagcc 300 cgcggacccc gagcacgcgc ctgacagccc ctgctggcccggcgcgcggc gtcgccaggc 360 cagctatggc ccccgacccg gtggccgccg agaccgcggctcagggacct accccgcgct 420 acttcacctg ggacgaggtg gcccagcgct cagggtgcgaggagcggtgg ctagtgatcg 480 accgtaaggt gtacaacatc agcgagttca cccgccggcatccagggggc tcccgggtca 540 tcagccacta cgccgggcag gatgccacgg tgagcgcagccaggcggggg cacaggagag 600 ggcgggaccg gaggctgagt gcaggggaga cagagttacgcactccgagc caaacaccga 660 ctaattcgga ggaaagcccg gaggcgcctg atcatacctgttgcccggtg attgggtgtc 720 ctgcggatgc gggatgaaaa ggcgggagag aggcctggagaagtggagtc tggggagtgg 780 ggatggaggc caacaacacg cacacacaaa caaagggtcccgcctccctg ccgtgcattc 840 catctgcagc cccgagcctc aggtctctgg gcggggacagaaccccgagc tgggtaggct 900 aggagggagg agagcaagga tgcaggccgc ctggggagggagggggtcag tggccagggg 960 agggagtcac atcctgtctc gatggctagg agaggcagcgcagccgcgtc tggacctagg 1020 tgccggtctc cactcgccag caggagcgga gagggagcaggaaaggagcc cattctcgag 1080 gatggggctg aaacgggaag cttggggaga ccgctgccttggggacccct gcgtcgtgtg 1140 aagactggag gacgcggaag ggacagcgct ggccggggagggcaagcggc cgctggcgta 1200 cataagggat tgggaatggc atacacttag cgaggacccccagagctgtt ctcgaatcgc 1260 ggggaggccc tgagccgcag gccagcgagg tcttcagctattccgcggag cggaccgctg 1320 tttacgctct ggggcggtag gcccttcgcg gggtcctgtcccttcttccc ttggtctcac 1380 tgcggggtcg gcgcgcgccc cagccccagg cctgctgcttccctttctag accacagccc 1440 tcagagctaa ggccccggcg cctctctgct gggttggagtcctggggact cagtcctagg 1500 gactcgaaag tcggggcgtt cccttcaccg cgtttcccccttggcggcca gaatggcgtc 1560 ccctcccctt gcatccccct ctgatcccgt gccctgcagcgtgatgccct ccactgtccc 1620 tatccactac cctggcgtcc cagagtgtgc cgcgggtcaccaggttccca taacgtcgca 1680 gcagagctta gacgctgcgg ggcgaagacc cgccccaccctctgacgcga ccagcctagt 1740 gggcgaggcc agagcttgcg cgggtcaacc agagtgaccactcgggagcc ctgactgcgg 1800 ccaagggcgc aggcgtgtcc cggcgcatgc gcagacgaaacaggcaccaa cgctggagct 1860 tcccgcagtg tgatttgggg ccggggatgc cgcggcggggacggcgattg gtccgtatgt 1920 gtggtgccac cggccgccgc tccgccccgg cccccgccccacacgccgca tcacttacag 1980 ggcccggggc tgccggacct gccaacgtga atcttatcgccatggacctt accttgcaca 2040 acccaaagta gctgccttgg ggcagggggt ggccagagtgcttagggaaa tgtggagccc 2100 tacccagaac aacggtggag ggaaagggaa gaaacgcagaagtgccccag ttcggacgta 2160 gggaagtctt cctcttcgtg gtttttggag aaccctagctaagagaggaa agggacttat 2220 tgaaagaccc gcaagaaggg acggaagtct catagccctgagaggtgaag ccagctggag 2280 ttgatgggtc gaatggggac ctagagaact tttctgtatctagaggtttg taaaatgcac 2340 caatcagtgc tctgtaaaaa cgcaccaatt ggcgctctgtagctagctag aggtttgtaa 2400 aatgagccaa tcagcaggac gtgggcaggg acaactaagacaataaaagc tggccacccc 2460 agccagctgc tgcaacccgc tccagttccc ttacaggctgtggaagcatt gttcttttgc 2520 tcgtcacact aaaccttgct gctgctcatt ctttgggtctgcaaagagtg ttattccttt 2580 aagagctata acagcgggaa ggtccacggc tccattcttgaagtcagtga gaccataccc 2640 gccggaagga accaacgccc gacacagccc cacccatctctcctgtttct cacctatact 2700 gaaattcttg ggcaaaagct gtctgtggac acacccaggggaaaggccag cccaggcagg 2760 tgtttcttag tggttcccct cagccaatgc ttcccattccttgatgcatc cttctaacta 2820 gagcagatgc tcggtgatct taaactgtgg acacctgggagcaccctcaa aaggcagctg 2880 ggcctaggga gatggcctgt gcttctgtgt caggagttggttccttcagg tgggcttgtg 2940 gtctcgctga cgtcaagaat gaagccatga accttcgcggtgagtgttac agctcttaca 3000 ggtggcgtgg acccaaagag tgagcagcag caagatttattgtgaagagc aaagaacaaa 3060 gcttccacag cgtggaaggg tacccgagca ggttgccgctgctggacgtt ggggggtgtg 3120 agggggagca gccttttttt ttcttttttt tttgagacggagtctccctg tcgcccaggc 3180 tggagtgcag tggcgcgatc tcggctcact gcaggctccgcccccccccc ggggttcacg 3240 ccattctcct gcctcagcct cccgagtagc tgggactacaggcgcccgct acctcgcccg 3300 gctaattttt tgtattttta gtagagacgg ggtttcactgtgttagccag gatggtctcg 3360 atctcctgac ctcgtgatcc acccgccttg gcctcccaaagtgctgggat tacaggcgtg 3420 agccaccgcg cccggccggg agcagctttt attcccttatttgtccctgc ccatgtcctg 3480 ctgatttgtc cattttatag agcactgatt ggtccattttacagggtgct gattggtcca 3540 ttttacctct agctagctaa agagcacgga ttggtgcattttacaaacct ctagctacag 3600 aaaagttctc caagtctgca ctcgacccag gaagtccatctggcttcacc tctcacttca 3660 acttgggtac agccttctgg cgggcaggag gatggcctttggtgcgaaca ctgccggagt 3720 ccagggggct ggctccctca cctttcatct tctcccggcacttgcaggat ccctttgtgg 3780 ccttccacat caacaagggc cttgtgaaga agtatatgaactctctcctg attggagaac 3840 tgtctccaga gcagcccagc tttgagccca ccaagaatgtaagaccctgt gtttgctatg 3900 tcgcaactat tggttgttga gggggacaga gagggggtggaaggagagtc tagatggaat 3960 cacagtcata gtaatcacag tcagtagtag ctctggggagtcttgaggtc cctgcttctc 4020 ttgcatagtc atgaggtcac aggcccaagg gagcatggctttgcaaccta tggctccccc 4080 aaggctgcca ctaccatggc tgccatcatt gttatcatcattgttatcat atgagcactt 4140 actatgcacc aagcataaac tcataactct tacacatttacagatgagat aacaggctca 4200 gggaggttaa gcaacacagc caaggatcac acagttagtaaatggcagag caaggactta 4260 gtcccctgaa ctcttaggca ctatcccatg gcacctcctcctgtcatcct cattgtcgtg 4320 gtatctttgc ctaggactgt ggacttccca cagctacctcagtgggaggt ccttgagcct 4380 gagagggccc ttgtctccag tagcattggg gtgcagatgagaagaataac agctcctctt 4440 cctcttctgc agaaagagct gacagatgag ttccgggagctgcgggccac agtggagcgg 4500 atggggctca tgaaggccaa ccatgtcttc ttcctgctgtacctgctgca catcttgctg 4560 ctggatggtg cagcctggct caccctttgg gtctttgggacgtccttttt gcccttcctc 4620 ctctgtgcgg tgctgctcag tgcagttcag gtgagagcctttggcttgtc aagtgcacag 4680 caatgctcag catccctg 4698 34 990 DNA Homosapiens 34 atggcccccg acccggtggc cgccgagacc gcggctcagg gacctaccccgcgctacttc 60 acctgggacg aggtggccca gcgctcaggg tgcgaggagc ggtggctagtgatcgaccgt 120 aaggtgtaca acatcagcga gttcacccgc cggcatccag ggggctcccgggtcatcagc 180 cactacgccg ggcaggatgc cacggatccc tttgtggcct tccacatcaacaagggcctt 240 gtgaagaagt atatgaactc tctcctgatt ggagaactgt ctccagagcagcccagcttt 300 gagcccacca agaataaaga gctgacagat gagttccggg agctgcgggccacagtggag 360 cggatggggc tcatgaaggc caaccatgtc ttcttcctgc tgtacctgctgcacatcttg 420 ctgctggatg gtgcagcctg gctcaccctt tgggtctttg ggacgtcctttttgcccttc 480 ctcctctgtg cggtgctgct cagtgcagtt caggcccagg ctggctggctgcagcatgac 540 tttgggcacc tgtcggtctt cagcacctca aagtggaacc atctgctacatcattttgtg 600 attggccacc tgaagggggc ccccgccagt tggtggaacc acatgcacttccagcaccat 660 gccaagccca actgcttccg caaagaccca gacatcaaca tgcatcccttcttctttgcc 720 ttggggaaga tcctctctgt ggagcttggg aaacagaaga aaaaatatatgccgtacaac 780 caccagcaca aatacttctt cctaattggg cccccagcct tgctgcctctctacttccag 840 tggtatattt tctattttgt tatccagcga cccccagcct tgctgcctctctacttccag 900 tggtatattt tctattttgt tatccagcga aagaagtggg tggacttggcctggatgatt 960 accttctacg tccgcttctt cctcacttat 990 35 960 DNA Homosapiens 35 ggcccggcgc gcggcgtcgc caggccagct atggcccccg acccggtggccgccgagacc 60 gcggctcagg gacctacccc gcgctacttc acctgggacg aggtggcccagcgctcaggg 120 tgcgaggagc ggtggctagt gatcgaccgt aaggtgtaca acatcagcgagttcacccgc 180 cggcatccag ggggctcccg ggtcatcagc cactacgccg ggcaggatgccacggatccc 240 tttgtggcct tccacatcaa caagggcctt gtgaagaagt atatgaactctctcctgatt 300 ggagaactgt ctccagagca gcccagcttt gagcccacca agaataaagagctgacagat 360 gagttccggg agctgcgggc cacagtggag cggatggggc tcatgaaggccaaccatgtc 420 ttcttcctgc tgtacctgct gcacatcttg ctgctggatg gtgcagcctggctcaccctt 480 tgggtctttg ggacgtcctt tttgcccttc ctcctctgtg cggtgctgctcagtgcagtt 540 caggcccagg ctggctggct gcagcatgac tttgggcacc tgtcggtcttcagcacctca 600 aagtggaacc atctgctaca tcattttgtg attggccacc tgaagggggcccccgccagt 660 tggtggaacc acatgcactt ccagcaccat gccaagccca actgcttccgcaaagaccca 720 gacatcaaca tgcatccctt cttctttgcc ttggggaaga tcctctctgtggagcttggg 780 aaacagaaga aaaaatatat gccgtacaac caccagcaca aatacttcttcctaattggg 840 cccccagcct tgctgcctct ctacttccag tggtatattt tctattttgttatccagcga 900 aagaagtggg tggacttggc ctggatcagc aaacaggaat acgatgaagccgggcttcca 960 36 473 DNA Homo sapiens 36 tccagcgaaa gaagtgggtggacttggcct ggatgattac cttctacgtc cgcttcttcc 60 tcacttatgt gccactattggggctgaaag ccttcctggg ccttttcttc atagtcaggt 120 tcctggaaag caactggtttgtgtgggtga cacagatgaa ccatattccc atgcacattg 180 atcatgaccg gaacatggactgggtttcca cccagctcct ggccacatgc aatgtccaca 240 agtctgcctt caatgactggttcagtggac acctcaactt ccagattgag caccatcttt 300 ttcccacgat gcctcgacacaattaccaca aagtggctcc cctggtgcag tccttgtgtg 360 ccaagcgtgg catagagtaccagtccaagc ccctgctgtc agccttcgcc gacatcatcc 420 actcactaaa ggagtcagggcagctctggc tagatgccta tcttcaccaa taa 473 37 449 DNA Homo sapiensmisc_feature (5)...(5) k = g or t/u at position 5 37 gaatkmttaccttctacgtc cgcttcttcc tcacttatgt gccactattg gggctgaaag 60 cttcctgggccttttcttca tagtcaggtt cctggaaagc aactggtttg tgtgggtgac 120 acagatgaaccatattccca tgcacattga tcatgaccgg aacatggact gggtttccac 180 ccagctccaggccacatgca atgtccacaa gtctgccttc aatgactggt tcagtggaca 240 cctcaacttccagattgagc accatctttt tcccacgatg cctcgacaca attaccacaa 300 agtggctcccctggtgcagt ccttgtgtgc caagcatggc atagagtacc agtccaagcc 360 cctgctgtcagccttcgccg acatcatcca ctcactaaag gagtcagggc agctctggct 420 agatgcctatcttcaccaat aacaacagc 449 38 465 PRT Homo sapiens VARIANT (432)...(432)Xaa = Unknown or other at position 432 38 Gln Gly Pro Thr Pro Arg TyrPhe Thr Trp Asp Glu Val Ala Gln Arg 1 5 10 15 Ser Gly Cys Glu Glu ArgTrp Leu Val Ile Asp Arg Lys Val Tyr Asn 20 25 30 Ile Ser Glu Phe Thr ArgArg His Pro Gly Gly Ser Arg Val Ile Ser 35 40 45 His Tyr Ala Gly Gln AspAla Thr Asp Pro Phe Val Ala Phe His Ile 50 55 60 Asn Lys Gly Leu Val LysLys Tyr Met Asn Ser Leu Leu Ile Gly Glu 65 70 75 80 Leu Ser Pro Glu GlnPro Ser Phe Glu Pro Thr Lys Asn Lys Glu Leu 85 90 95 Thr Asp Glu Phe ArgGlu Leu Arg Ala Thr Val Glu Arg Met Gly Leu 100 105 110 Met Lys Ala AsnHis Val Phe Phe Leu Leu Tyr Leu Leu His Ile Leu 115 120 125 Leu Leu AspGly Ala Ala Trp Leu Thr Leu Trp Val Phe Gly Thr Ser 130 135 140 Phe LeuPro Phe Leu Leu Cys Ala Val Leu Leu Ser Ala Val Gln Ala 145 150 155 160Gln Ala Gly Trp Leu Gln His Asp Tyr Gly His Leu Ser Val Tyr Arg 165 170175 Lys Pro Lys Trp Asn His Leu Val His Lys Phe Val Ile Gly His Leu 180185 190 Lys Gly Ala Ser Ala Asn Trp Trp Asn His Arg His Phe Gln His His195 200 205 Ala Lys Pro Asn Ile Phe His Lys Asp Pro Asp Val Asn Met LeuHis 210 215 220 Val Phe Val Leu Gly Glu Trp Gln Pro Ile Glu Tyr Gly LysLys Lys 225 230 235 240 Leu Lys Tyr Leu Pro Tyr Asn His Gln His Glu TyrPhe Phe Leu Ile 245 250 255 Gly Pro Pro Leu Leu Ile Pro Met Tyr Phe GlnTyr Gln Ile Ile Met 260 265 270 Thr Met Ile Val His Lys Asn Trp Val AspLeu Ala Trp Ala Val Ser 275 280 285 Tyr Tyr Ile Arg Phe Phe Ile Thr TyrIle Pro Phe Tyr Gly Ile Leu 290 295 300 Gly Ala Leu Leu Phe Leu Asn PheIle Arg Phe Leu Glu Ser His Trp 305 310 315 320 Phe Val Trp Val Thr GlnMet Asn His Ile Val Met Glu Ile Asp Gln 325 330 335 Glu Ala Tyr Arg AspTrp Phe Ser Ser Gln Leu Thr Ala Thr Cys Asn 340 345 350 Val Glu Gln SerPhe Phe Asn Asp Trp Phe Ser Gly His Leu Asn Phe 355 360 365 Gln Ile GluHis His Leu Phe Pro Thr Met Pro Arg His Asn Leu His 370 375 380 Lys IleAla Pro Leu Val Lys Ser Leu Cys Ala Lys His Gly Ile Glu 385 390 395 400Tyr Gln Glu Lys Pro Leu Leu Arg Ala Leu Leu Asp Ile Ile Arg Ser 405 410415 Leu Lys Lys Ser Gly Lys Leu Trp Leu Asp Ala Tyr Leu His Lys Xaa 420425 430 Ser His Ser Pro Arg Asp Thr Val Gly Lys Gly Cys Arg Trp Gly Asp435 440 445 Gly Gln Arg Asn Asp Gly Leu Leu Phe Xaa Gly Val Ser Glu ArgLeu 450 455 460 Val 465 39 360 PRT Homo sapiens 39 Met Ala Pro Asp ProVal Ala Ala Glu Thr Ala Ala Gln Gly Pro Thr 1 5 10 15 Pro Arg Tyr PheThr Trp Asp Glu Val Ala Gln Arg Ser Gly Cys Glu 20 25 30 Glu Arg Trp LeuVal Ile Asp Arg Lys Val Tyr Asn Ile Ser Glu Phe 35 40 45 Thr Arg Arg HisPro Gly Gly Ser Arg Val Ile Ser His Tyr Ala Gly 50 55 60 Gln Asp Ala ThrAsp Pro Phe Val Ala Phe His Ile Asn Lys Gly Leu 65 70 75 80 Val Lys LysTyr Met Asn Ser Leu Leu Ile Gly Glu Leu Ser Pro Glu 85 90 95 Gln Pro SerPhe Glu Pro Thr Lys Asn Lys Glu Leu Thr Asp Glu Phe 100 105 110 Arg GluLeu Arg Ala Thr Val Glu Arg Met Gly Leu Met Lys Ala Asn 115 120 125 HisVal Phe Phe Leu Leu Tyr Leu Leu His Ile Leu Leu Leu Asp Gly 130 135 140Ala Ala Trp Leu Thr Leu Trp Val Phe Gly Thr Ser Phe Leu Pro Phe 145 150155 160 Leu Leu Cys Ala Val Leu Leu Ser Ala Val Gln Ala Gln Ala Gly Trp165 170 175 Leu Gln His Asp Phe Gly His Leu Ser Val Phe Ser Thr Ser LysTrp 180 185 190 Asn His Leu Leu His His Phe Val Ile Gly His Leu Lys GlyAla Pro 195 200 205 Ala Ser Trp Trp Asn His Met His Phe Gln His His AlaLys Pro Asn 210 215 220 Cys Phe Arg Lys Asp Pro Asp Ile Asn Met His ProPhe Phe Phe Ala 225 230 235 240 Leu Gly Lys Ile Leu Ser Val Glu Leu GlyLys Gln Lys Lys Lys Tyr 245 250 255 Met Pro Tyr Asn His Gln His Lys TyrPhe Phe Leu Ile Gly Pro Pro 260 265 270 Ala Leu Leu Pro Leu Tyr Phe GlnTrp Tyr Ile Phe Tyr Phe Val Ile 275 280 285 Gln Arg Lys Lys Trp Val AspLeu Ala Trp Met Ile Thr Phe Tyr Val 290 295 300 Arg Phe Phe Leu Thr TyrVal Pro Leu Leu Gly Leu Lys Ala Phe Leu 305 310 315 320 Gly Leu Phe PheIle Val Arg Phe Leu Glu Ser Asn Trp Phe Val Trp 325 330 335 Val Thr GlnMet Asn His Ile Pro Met His Ile Asp His Asp Arg Asn 340 345 350 Met AspTrp Val Ser Thr Gln Leu 355 360 40 347 PRT Homo sapiens VARIANT(251)...(251) Xaa = Unknown or other at position 251 40 Gln Gly Pro ThrPro Arg Tyr Phe Thr Trp Asp Glu Val Ala Gln Arg 1 5 10 15 Ser Gly CysGlu Glu Arg Trp Leu Val Ile Asp Arg Lys Val Tyr Asn 20 25 30 Ile Ser GluPhe Thr Arg Arg His Pro Gly Gly Ser Arg Val Ile Ser 35 40 45 His Tyr AlaGly Gln Asp Ala Thr Asp Pro Phe Val Ala Phe His Ile 50 55 60 Asn Lys GlyLeu Val Lys Lys Tyr Met Asn Ser Leu Leu Ile Gly Glu 65 70 75 80 Leu SerPro Glu Gln Pro Ser Phe Glu Pro Thr Lys Asn Lys Glu Leu 85 90 95 Thr AspGlu Phe Arg Glu Leu Arg Ala Thr Val Glu Arg Met Gly Leu 100 105 110 MetLys Ala Asn His Val Phe Phe Leu Leu Tyr Leu Leu His Ile Leu 115 120 125Leu Leu Asp Gly Ala Ala Trp Leu Thr Leu Trp Val Phe Gly Thr Ser 130 135140 Phe Leu Pro Phe Leu Leu Cys Ala Val Leu Leu Ser Ala Val Gln Ala 145150 155 160 Gln Ala Gly Trp Leu Gln His Asp Gly His Leu Ser Val Phe SerThr 165 170 175 Ser Lys Trp Asn His Leu Leu His His Phe Val Ile Gly HisLeu Lys 180 185 190 Gly Ala Pro Ala Ser Trp Trp Asn His Met His Phe GlnHis His Ala 195 200 205 Lys Pro Asn Cys Phe Arg Lys Asp Pro Asp Ile AsnMet His Pro Phe 210 215 220 Phe Phe Ala Leu Gly Lys Ile Leu Ser Val GluLeu Gly Lys Gln Lys 225 230 235 240 Lys Lys Tyr Met Pro Tyr Asn His GlnHis Xaa Tyr Phe Phe Leu Ile 245 250 255 Gly Pro Pro Ala Leu Leu Pro LeuTyr Phe Gln Trp Tyr Ile Phe Tyr 260 265 270 Phe Val Ile Gln Arg Lys LysTrp Val Asp Leu Ala Trp Ile Ser Lys 275 280 285 Gln Glu Tyr Asp Glu AlaGly Leu Pro Leu Ser Thr Ala Asn Ala Ser 290 295 300 Lys Arg Asp Leu ProArg Ala Thr Ser Pro Gly Thr Arg Trp Pro Ser 305 310 315 320 Ala Gln GlyAla Arg Ser Gly Gly Xaa Xaa Ser Thr Val Arg Cys Thr 325 330 335 Thr SerAla Ser Ser Pro Ala Gly Ile Gln Gly 340 345 41 458 PRT Homo sapiensVARIANT (458)...(458) Xaa = Unknown or other at position 458 41 Met AlaAla Ala Pro Ser Val Arg Thr Phe Thr Arg Ala Glu Val Leu 1 5 10 15 AsnAla Glu Ala Leu Asn Glu Gly Lys Lys Asp Ala Glu Ala Pro Phe 20 25 30 LeuMet Ile Ile Asp Asn Lys Val Tyr Asp Val Arg Glu Phe Val Pro 35 40 45 AspHis Pro Gly Gly Ser Val Ile Leu Thr His Val Gly Lys Asp Gly 50 55 60 ThrAsp Val Phe Asp Thr Phe His Pro Glu Ala Ala Trp Glu Thr Leu 65 70 75 80Ala Asn Phe Tyr Val Gly Asp Ile Asp Glu Ser Asp Arg Asp Ile Lys 85 90 95Asn Asp Asp Phe Ala Ala Glu Val Arg Lys Leu Arg Thr Leu Phe Gln 100 105110 Ser Leu Gly Tyr Tyr Asp Ser Ser Lys Ala Tyr Tyr Ala Phe Lys Val 115120 125 Ser Phe Asn Leu Cys Ile Trp Gly Leu Ser Thr Val Ile Val Ala Lys130 135 140 Trp Gly Gln Thr Ser Thr Leu Ala Asn Val Leu Ser Ala Ala LeuLeu 145 150 155 160 Gly Leu Phe Trp Gln Gln Cys Gly Trp Leu Ala His AspPhe Leu His 165 170 175 His Gln Val Phe Gln Asp Arg Phe Trp Gly Asp LeuPhe Gly Ala Phe 180 185 190 Leu Gly Gly Val Cys Gln Gly Phe Ser Ser SerTrp Trp Lys Asp Lys 195 200 205 His Asn Thr His His Ala Ala Pro Asn ValHis Gly Glu Asp Pro Asp 210 215 220 Ile Asp Thr His Pro Leu Leu Thr TrpSer Glu His Ala Leu Glu Met 225 230 235 240 Phe Ser Asp Val Pro Asp GluGlu Leu Thr Arg Met Trp Ser Arg Phe 245 250 255 Met Val Leu Asn Gln ThrTrp Phe Tyr Phe Pro Ile Leu Ser Phe Ala 260 265 270 Arg Leu Ser Trp CysLeu Gln Ser Ile Leu Phe Val Leu Pro Asn Gly 275 280 285 Gln Ala His LysPro Ser Gly Ala Arg Val Pro Ile Ser Leu Val Glu 290 295 300 Gln Leu SerLeu Ala Met His Trp Thr Trp Tyr Leu Ala Thr Met Phe 305 310 315 320 LeuPhe Ile Lys Asp Pro Val Asn Met Leu Val Tyr Phe Leu Val Ser 325 330 335Gln Ala Val Cys Gly Asn Leu Leu Ala Ile Val Phe Ser Leu Asn His 340 345350 Asn Gly Met Pro Val Ile Ser Lys Glu Glu Ala Val Asp Met Asp Phe 355360 365 Phe Thr Lys Gln Ile Ile Thr Gly Arg Asp Val His Pro Gly Leu Phe370 375 380 Ala Asn Trp Phe Thr Gly Gly Leu Asn Tyr Gln Ile Glu His HisLeu 385 390 395 400 Phe Pro Ser Met Pro Arg His Asn Phe Ser Lys Ile GlnPro Ala Val 405 410 415 Glu Thr Leu Cys Lys Lys Tyr Asn Val Arg Tyr HisThr Thr Gly Met 420 425 430 Ile Glu Gly Thr Ala Glu Val Phe Ser Arg LeuAsn Glu Val Ser Lys 435 440 445 Ala Ala Ser Lys Met Gly Lys Ala Gln Xaa450 455 42 444 PRT Homo sapiens VARIANT (444)...(444) Xaa = Unknown orother at position 444 42 Met Ala Pro Asp Pro Val Ala Ala Glu Thr Ala AlaGln Gly Pro Thr 1 5 10 15 Pro Arg Tyr Phe Thr Trp Asp Glu Val Ala GlnArg Ser Gly Cys Glu 20 25 30 Glu Arg Trp Leu Val Ile Asp Arg Lys Val TyrAsn Ile Ser Glu Phe 35 40 45 Thr Arg Arg His Pro Gly Gly Ser Arg Val IleSer His Tyr Ala Gly 50 55 60 Gln Asp Ala Thr Asp Pro Phe Val Ala Phe HisIle Asn Lys Gly Leu 65 70 75 80 Val Lys Lys Tyr Met Asn Ser Leu Leu IleGly Glu Leu Ser Pro Glu 85 90 95 Gln Pro Ser Phe Glu Pro Thr Lys Asn LysGlu Leu Thr Asp Glu Phe 100 105 110 Glu Leu Arg Ala Thr Val Glu Arg MetGly Leu Met Lys Ala Asn His 115 120 125 Val Phe Phe Leu Leu Tyr Leu LeuHis Ile Leu Leu Leu Asp Gly Ala 130 135 140 Ala Trp Leu Thr Leu Trp ValPhe Gly Thr Ser Phe Leu Pro Phe Leu 145 150 155 160 Leu Cys Ala Val LeuLeu Ser Ala Val Gln Ala Gln Ala Gly Trp Leu 165 170 175 Gln His Asp PheGly His Leu Ser Val Phe Ser Thr Ser Lys Trp Asn 180 185 190 His Leu LeuHis His Phe Val Ile Gly His Leu Lys Gly Ala Pro Ala 195 200 205 Ser TrpTrp Asn His Met His Phe Gln His His Ala Lys Pro Asn Cys 210 215 220 PheArg Lys Asp Pro Asp Ile Asn Met His Pro Phe Phe Phe Ala Leu 225 230 235240 Gly Lys Ile Leu Ser Val Glu Leu Gly Lys Gln Lys Lys Lys Tyr Met 245250 255 Pro Tyr Asn His Gln His Lys Tyr Phe Phe Leu Ile Gly Pro Pro Ala260 265 270 Leu Leu Pro Leu Tyr Phe Gln Trp Tyr Ile Phe Tyr Phe Val IleGln 275 280 285 Arg Lys Lys Trp Val Asp Leu Ala Trp Met Ile Thr Phe TyrVal Arg 290 295 300 Phe Phe Leu Thr Tyr Val Pro Leu Leu Gly Leu Lys AlaPhe Leu Gly 305 310 315 320 Leu Phe Phe Ile Val Arg Phe Leu Glu Ser AsnTrp Phe Val Trp Val 325 330 335 Thr Gln Met Asn His Ile Pro Met His IleAsp His Asp Arg Asn Met 340 345 350 Asp Trp Val Ser Thr Gln Leu Leu AlaThr Cys Asn Val His Lys Ser 355 360 365 Ala Phe Asn Asp Trp Phe Ser GlyHis Leu Asn Phe Gln Ile Glu His 370 375 380 His Leu Phe Pro Thr Met ProArg His Asn Tyr His Lys Val Ala Pro 385 390 395 400 Leu Val Gln Ser LeuCys Ala Lys Arg Gly Ile Glu Tyr Gln Ser Lys 405 410 415 Pro Leu Leu SerAla Phe Ala Asp Ile Ile His Ser Leu Lys Glu Ser 420 425 430 Gly Gln LeuTrp Leu Asp Ala Tyr Leu His Gln Xaa 435 440 43 21 DNA ArtificialSequence Vector Primer RO329 43 cagaccaact ggtaatggta g 21 44 29 DNAArtificial Sequence Vector Primer RO384 44 tcaggcccaa gctggatggctgcaacatg 29 45 21 DNA Artificial Sequence Vector Primer RO328 45ctcctggagc ccgtcagtat c 21 46 29 DNA Artificial Sequence Vector PrimerRO388 46 atggtgggga agaggtggtg ctcaatctg 29 47 31 DNA ArtificialSequence Primer RO430 47 gtggctgttg ttattggtga agataggcat c 31 48 20 DNAArtificial Sequence Primer RO526 48 catggccccc gacccggtgg 20 49 20 DNAArtificial Sequence Primer RO527 49 gcggccaccg ggtcgggggc 20 50 34 DNAArtificial Sequence Primer RO512 50 gattgggtgc catggggatg cgggatgaaaaggc 34 51 18 DNA Artificial Sequence Primer RO5 51 gaaacagcta tgaccatg18 52 34 DNA Artificial Sequence Primer RO580 52 tcctgcgaat tcaccatgaaaaggcgggag agag 34 53 35 DNA Artificial Sequence Primer RO578 53catggctagg agaggcagcg cagccgcgtc tggac 35 54 35 DNA Artificial SequencePrimer RO579 54 ctaggtccag acgcggctgc gctgcctctc ctagc 35

1. An isolated nucleotide sequence corresponding to or complementary toat least about 50% of the nucleotide sequence represented by SEQ ID NO:1(FIG. 12).
 2. The isolated nucleotide sequence of claim 1 wherein saidsequence is represented by SEQ ID NO:1.
 3. The isolated nucleotidesequence of claims 1 or 2 wherein said sequence encodes a functionallyactive desaturase which utilizes a polyunsaturated fatty acid as asubstrate.
 4. The nucleotide sequence of claim 1 wherein said sequenceis derived from a mammal.
 5. The nucleotide sequence of claim 4 whereinsaid Sequence is derived from a human.
 6. A purified protein encoded bysaid nucleotide sequence of claims 1 or
 2. 7. A purified polypeptidewhich desaturates polyunsaturated fatty acids at carbon 5 and has atleast about 50% amino acid similarity to the amino acid sequence of saidpurified protein of claim
 6. 8. A method of producing a humanΔ5-desaturase comprising the steps of: a) isolating said nucleotidesequence represented by SEQ ID NO:1 (FIG. 12); b) constructing a vectorcomprising: i) said isolated nucleotide sequence operably linked to ii)a promoter; c) introducing said vector into a host cell under time andconditions sufficient for expression of said human Δ5-desaturase.
 9. Themethod of claim 8 wherein said host cell is a eukaryotic cell or aprokaryotic cell.
 10. The method of claim 9 wherein said prokaryoticcell is selected from the group consisting of E. coli, cyanobacteria,and B. subtilis.
 11. The method of claim 9 wherein said eukaryotic cellis selected from the group consisting of a mammalian cell, an insectcell, a plant cell and a fungal cell.
 12. The method of claim 11 whereinsaid fungal cell is a yeast cell.
 13. The method of claim 12 whereinsaid yeast cell is selected from the group consisting of Saccharomycescerevisiae, Saccharomyces carlsbergensis, Candida spp., Lipomycesstarkey, Yarrowia lipolytica, Kluyveromyces spp., Hansenula spp.,Trichoderma spp. and Pichia spp.
 14. The method of claim 13 wherein saidyeast cell is Saccharomyces cerevisiae.
 15. A vector comprising: a) anucleotide sequence as represented by SEQ ID NO:1 (FIG. 12) operablylinked to b) a promoter.
 16. A host cell comprising said vector of claim15.
 17. The host cell of claim 16, wherein said host cell is aeukaryotic cell or a prokaryotic cell.
 18. The host cell of claim 17wherein said prokaryotic cell is selected from the group consisting ofE. coli, Cyanobacteria, and B. subtilis.
 19. The host cell of claim 17wherein said eukaryotic cell is selected from the group consisting of amammalian cell, an insect cell, a plant cell and a fungal cell.
 20. Thehost cell of claim 19 wherein said fungal cell is a yeast cell.
 21. Thehost cell of claim 20 wherein said yeast cell is selected from the groupconsisting of Saccharomyces cerevisiae, Saccharomyces carlsbergensis,Candida spp., Lipomyces starkey, Yarrowia lipolytica, Kluyveromycesspp., Hansenula spp., Trichoderma spp. and Pichia spp.
 22. The host cellof claim 21 wherein said host cell is Saccharomyces cerevisiae.
 23. Aplant cell, plant or plant tissue comprising said vector of claim 15,wherein expression of said nucleotide sequence of said vector results inproduction of a polyunsaturated fatty acid by said plant cell or tissue.24. The plant cell, plant or plant tissue of claim 23 wherein saidpolyunsaturated fatty acid is AA or EPA.
 25. One or more plant oils oracids expressed by said plant cell, plant or plant tissue of claim 23.26. A transgenic plant comprising said vector of claim 15, whereinexpression of said nucleotide sequence of said vector results inproduction of a polyunsaturated fatty acid in seeds of said transgenicplant.
 27. A mammalian cell comprising said vector of claim 15, whereinexpression of said nucleotide sequence of said vector results inproduction of altered levels of AA or EPA when said cell is grown in aculture media comprising a fatty acid selected from the group consistingof an essential fatty acid, LA and ALA.
 28. A transgenic, non-humanmammal whose genome comprises a DNA sequence encoding a humanΔ5-desaturase operably linked to a promoter.
 29. The transgenic,non-human mammal of claim 28, wherein said DNA sequence is representedby SEQ ID NO:1 (FIG. 12).
 30. A fluid produced by said transgenic,non-human mammal of claim 29 wherein said fluid comprises a detectablelevel of at least human Δ5-desaturase.
 31. A method for producing apolyunsaturated fatty acid comprising the steps of: a) isolating saidnucleotide sequence represented by SEQ ID NO:1 (FIG. 12); b)constructing a vector comprising said isolated nucleotide sequence; c)introducing said vector into a host cell under time and conditionssufficient for expression of said human Δ5-desaturase enzyme; and d)exposing said expressed human Δ5-desaturase enzyme to a substratepolyunsaturated fatty acid in order to convert said substrate to aproduct polyunsaturated fatty acid.
 32. The method according to claim31, wherein said substrate polyunsaturated fatty acid is DGLA or 20:4n−3and said product polyunsaturated fatty acid is AA or EPA, respectively.33. The method according to claim 31 further comprising the step ofexposing said product polyunsaturated fatty acid to an elongase in orderto convert said product polyunsaturated fatty acid to anotherpolyunsaturated fatty acid.
 34. The method according to claim 33 whereinsaid product polyunsaturated fatty acid is AA or EPA and said anotherpolyunsaturated fatty acid is adrenic acid or (n−3)-docosapentaenoicacid, respectively.
 35. The method of claim 33 further comprising thesteps of exposing said another polyunsaturated fatty acid to anadditional desaturase in order to convert said another polyunsaturatedfatty acid to a final polyunsaturated fatty acid.
 36. The method ofclaim 35 wherein said final polyunsaturated fatty acid is(n−6)-docosapentaenoic acid or docosahexaenoic (DHA) acid.
 37. Anutritional composition comprising at least one polyunsaturated fattyacid selected from the group consisting of said product polyunsaturatedfatty acid produced according to the method of claim 31, said anotherpolyunsaturated fatty acid produced according to the method of claim 33,and said final polyunsaturated fatty acid produced according to themethod of claim
 35. 38. The nutritional composition of claim 37 whereinsaid product polyunsaturated fatty acid is AA or EPA.
 39. Thenutritional composition of claim 37 wherein said another polyunsaturatedfatty acid is adrenic acid or (n−3)-docosapentaenoic acid.
 40. Thenutritional composition of claim 37 wherein said final polyunsaturatedfatty acid is DHA.
 41. The nutritional composition of claim 37 whereinsaid nutritional composition is selected from the group consisting of aninfant formula, a dietary supplement and a dietary substitute.
 42. Thenutritional composition of claim 41 wherein said nutritional compositionis administered to a human or an animal.
 43. The nutritional compositionof claim 42 wherein said nutritional composition is administeredenterally or parenterally.
 44. The nutritional composition of claim 41wherein said nutritional composition further comprises at least onemacronutrient selected from the group consisting of coconut oil, soyoil, canola oil, monoglycerides, diglycerides, glucose, edible lactose,electrodialysed whey, electrodialysed skim milk, milk whey, soy protein,and protein hydrolysates.
 45. The nutritional composition of claim 44wherein said nutritional composition further comprises at least onevitamin selected from the group consisting of Vitamins A, C, D, E, and Bcomplex and at least one mineral selected from the group consisting ofcalcium magnesium, zinc, manganese, sodium, potassium, phosphorus,copper, chloride, iodine, selenium and iron.
 46. A pharmaceuticalcomposition comprising 1) at least one polyunsaturated fatty acidselected from the group consisting of said product polyunsaturated fattyacid produced according to the method of claim 31, said anotherpolyunsaturated fatty acid produced according to the method of claim 33,and said final polyunsaturated fatty acid produced according to themethod of claim 35 and 2) a pharmaceutically acceptable carrier.
 47. Thepharmaceutical composition of claim 46 wherein said pharmaceuticalcomposition is administered to a human or an animal.
 48. Thepharmaceutical composition of claim 46 wherein said pharmaceuticalcomposition further comprises an element selected from the groupconsisting of a vitamin, a mineral, a carbohydrate, an amino acid, afree fatty acid, a phospholipid, an antioxidant, and a phenoliccompound.
 49. An animal feed comprising at least one polyunsaturatedfatty acid selected from the group consisting of said productpolyunsaturated fatty acid produced according to the method of claim 31,said another polyunsaturated fatty acid produced according to the methodof claim 33 and said final polyunsaturated fatty acid produced accordingto the method of claim
 35. 50. The animal feed of claim 49 wherein saidproduct polyunsaturated fatty acid is AA or EPA.
 51. The animal feed ofclaim 49 wherein said another polyunsaturated fatty acid is adrenic acidor (n−3)-docosapentaenoic acid.
 52. The animal feed of claim 49 whereinsaid final polyunsaturated fatty acid is (n−6)-docosapentaenoic acid orDHA.
 53. A cosmetic comprising a polyunsaturated fatty acid selectedfrom the group consisting of said product polyunsaturated fatty acidproduced according to the method of claim 31, said anotherpolyunsaturated fatty acid produced according to the method of claim 33and said final polyunsaturated fatty acid produced according to themethod of claim
 35. 54. A method of preventing or treating a conditioncaused by insufficient intake of polyunsaturated fatty acids comprisingadministering to said patient said nutritional composition of claim 37in an amount sufficient to effect said treatment.